Salts of potassium atp channel openers and uses thereof

ABSTRACT

Provided are immediate or prolonged administration of certain salts of K ATP  channel openers such as diazoxide to a subject to achieve novel pharmacodynamic, pharmacokinetic, therapeutic, physiological, metabolic and compositional outcomes in the treatment of diseases or conditions involving K ATP  channels. Also provided are pharmaceutical formulations, methods of administration and dosing of the salts that achieve these outcomes and reduce the incidence of adverse effects in treated individuals. Further provided are method of co-administering the salts with other drugs to treat diseases of humans and animals.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of PCT International application serial number PCT/US2013/027676, filed on Feb. 25, 2013, which claims priority to U.S. Ser. No. 61/603,923, filed on Feb. 27, 2012, each of which is hereby incorporated by reference in its entirety.

FIELD OF THE INVENTION

The present invention relates to salts of potassium ATP (K_(ATP)) channel openers, methods of preparing such salts, and methods of use thereof for treatment of a variety of diseases and conditions, including for example, type 1 and type 2 diabetes, hypertension, dyslipidemia, nonalcoholic steatohepatitis, pulmonary hypertension, myocardial infarction and arrhythmias following myocardical infarction and poly-cystic ovarian syndrome.

BACKGROUND OF THE INVENTION

The following description of the background of the invention is provided as an aid in understanding the invention and is not admitted to describe or constitute prior art to the invention.

ATP-sensitive potassium (K_(ATP)) channels play important roles in a variety of tissues by coupling cellular metabolism to electrical activity. The K_(ATP) channel has been identified as an octameric complex of two unrelated proteins, which assemble in a 4:4 stoichiometry. The first is a pore forming subunit, Kir6.x, which forms an inwardly rectifying K⁺ channel; the second is an ABC (ATP binding cassette) transporter, also known as the sulfonylurea receptor (SURx) (Babenko et al., Annu. Rev. Physiol., 60:667-687 (1998)). The Kir6.x pore forming subunit is common for many types of K_(ATP) channels, and has two putative transmembrane domains (identified as TM1 and TM2), which are linked by a pore loop (H5). The subunit that comprises the SUR receptor includes multiple membrane-spanning domains and two nucleotide-binding folds.

According to their tissue localization, K_(ATP) channels exist in different isoforms or subspecies resulting from the assembly of the SUR and Kir subunits in multiple combinations. The combination of the SUR1 with the Kir6.2 subunits (SUR1/Kir6.2) typically forms the adipocyte and pancreatic β-cell type K_(ATP) channels, whereas the SUR2A/Kir6.2 and the SUR2B/Kir6.2 or Kir6.1 combinations typically form the cardiac type and the smooth muscle type K_(ATP) channels, respectively (Babenko et al., Annu. Rev. Physiol., 60:667-687 (1998)). There is also evidence that the channel may include Kir2.x subunits. This class of potassium channels are inhibited by intracellular ATP and activated by intracellular nucleoside diphosphates. Such K_(ATP) channels link the metabolic status of the cells to the plasma membrane potential and in this way play a key role in regulating cellular activity. In most excitatory cells, K_(ATP) channels are closed under normal physiological conditions and open when the tissue is metabolically compromised (e.g. when the (ATP:ADP) ratio falls). This promotes K⁺ efflux and cell hyperpolarization, thereby preventing voltage-dependent Ca²⁺ channels (VDCCs) from opening. (Prog. Res Research, (2001) 31:77-80).

Potassium channel openers (PCOs or KCOs; also referred to as channel activators or channel agonists), are a structurally diverse group of compounds with no apparent common pharmacophore linking their ability to antagonize the inhibition of K_(ATP) channels by intracellular nucleotides. Diazoxide is a PCO that stimulates K_(ATP) channels in pancreatic β-cells (see Trube et al., Pfluegers Arch Eur J Physiol, 407, 493-99 (1986)). Pinacidil and chromakalim are PCOs that activate sarcolemmal potassium channels (see Escande et al., Biochem Biophys Res Commun, 154, 620-625 (1988); Babenko et al., J Biol Chem, 275(2), 717-720 (2000)). Responsiveness to diazoxide has been shown to reside in the 6^(th) through 11^(th) predicted transmembrane domains (TMD6-11) and the first nucleotide-binding fold (NBF1) of the SUR1 subunit.

Diazoxide, which is a nondiuretic benzothiadiazine derivative having the formula 7-chloro-3-methyl-2H-1,2,4-benzothiadiazine 1.1-dioxide (empirical formula C₈H₇ClN₂O₂S), is commercialized in three distinct formulations to treat two different disease indications: (1) hypertensive emergencies and (2) hyperinsulinemic hypoglycemic conditions. Hypertensive emergencies are treated with Hyperstat IV, an aqueous formulation of diazoxide for intravenous use, adjusted to pH 11.6 with sodium hydroxide. Hyperstat IV is administered as a bolus dose into a peripheral vein to treat malignant hypertension or sulfonylurea overdose. In these uses, diazoxide acts to open potassium channels in vascular smooth muscle and pancreatic beta-cells, stabilizing the membrane potential at the resting level, resulting in vascular smooth muscle relaxation and suppression of insulin release, respectively.

Hyperinsulinemic hypoglycemic conditions are treated with Proglycem®, an oral pharmaceutical version of diazoxide useful for administration to infants, children and adults. It is available as a chocolate mint flavored oral suspension, which includes 7.25% alcohol, sorbitol, chocolate cream flavor, propylene glycol, magnesium aluminum silicate, carboxymethylcellulose sodium, mint flavor, sodium benzoate, methylparaben, hydrochloric acid to adjust the pH, poloxamer 188, propylparaben and water. Diazoxide is also available as a capsule with 50 or 100 mg of diazoxide including lactose and magnesium stearate. In these uses, diazoxide activated K_(ATP) channels in insulin secreting cells thereby blunting the hypersecreting conditions.

Myocardial remodeling late after infarction is associated with increased incidence of fatal arrhythmias. Heterogeneous prolongation of the action potential in the surviving myocardium is one of the predominant causes. Sarcolemmal ATP-dependent potassium (K_(ATP)) channels are important metabolic sensors regulating electrical activity of cardiomyocytes and are capable of considerably shortening the action potential. Tavares et al. (Expression and function of ATP-dependent potassium channels in late post-infarction remodeling, J Mol Cell Cardiol 42:1016-1025 (2007)) studied the effect of diazoxide on late post infarction remodeling in rats. Cardiomyocytes were obtained from the infarct border zone, the septum and the right ventricle of rat hearts 10 weeks after coronary occlusion when rats developed signs of heart failure. Expression of the conductance subunit Kir6.1, but not Kir6.2, and of all SUR regulatory subunits was increased up to 3-fold in cardiomyocytes from the infarct border zone. Concomitantly, there was a prominent response of the K_(ATP) current to diazoxide that was not detectable in control cardiomyocytes. The action potential was prolonged in cardiomyocytes from the infarct border zone (74 ms) relative to sham (41 ms). However, activation of the K_(ATP) channels by diazoxide reduced action potential duration to 42 ms. In myocytes of the septum and right ventricle, expression of channel subunits, duration of action potential, and sensitivity to diazoxide were only slightly increased relative to shams. The authors suggested that drugs selectively activating diazoxide-sensitive sarcolemmal K_(ATP) channels should be considered in the prevention of arrhythmias in post-infarction heart failure.

Schwartz et al. (Cardioprotection by multiple preconditioning cycles does not require mitochondrial KATP channels in pigs, Am J Physiol Heart Circ Physiol 283:H1538-H1544 (2002)) studied the effects of diazoxide preconditioning on infarct size in pigs. Diazoxide was administered 3.5 mg/kg, 1 ml/min IV to in barbital-anesthetized open-chest pigs subjected to 30 min of complete occlusion of the left anterior descending coronary artery and 3 h of reflow. Infarct size (percentage of the area at risk) after 30 min of ischemia in controls was 35.1±9.9% (n=7). Diazoxide infusion significantly limited infarct size (14.6±7.4%, n=7). Similar results have been demonstrated in rat and rabbit models.

Diazoxide administered either as an IV bolus or orally has been used to treat pulmonary hypertension. For example, Chan et al. (Reversibility of primary pulmonary hypertension during six years of treatment with oral diazoxide, Br Heart J57(2):207-209 (1987)) reported the successful treatment of a 32 year old woman with pulmonary hypertension. Her symptoms resolved completely with oral diazoxide and the pulmonary arterial pressure was reduced to normal levels over a period of six years. When diazoxide was discontinued on two separate occasions pulmonary hypertension recurred. Squarcia et al. (Primary pulmonary hypertension in childhood: familial aspects, Pediatr Med Chir 3(6):467-472 (1981)) suggested that among alternative vasodilators available for the experimental treatment of pulmonary hypertension diazoxide appears to have some advantages because it reduces not only pulmonary arteriolar resistance, but also pulmonary artery pressure, without producing tachycardia.

Honey et al. (Clinical and hemodynamic effects of diazoxide in primary pulmonary hypertension, Thorax 35(4):269-276 (1980)) studied the effects of IV and oral diazoxide on primary pulmonary hypertension. In their study diazoxide was injected into the pulmonary artery in nine patients with primary pulmonary hypertension. There was no significant change in pulmonary artery pressure, which fell by more than 10 mmHg in only two patients. The pulmonary blood flow increased in all patients as a result of a fall in pulmonary vascular resistance (by 4 to 17 units). Systematic vascular resistance also fell as expected in all patients. Oral diazoxide was given to seven patients, two of whom showed sustained clinical improvement while remaining on treatment (400 to 600 mg daily). Five patients were unable to tolerate the drug, because of nausea and sickness (two), peripheral edema requiring large doses of diuretics (four), diabetes (three), and postural hypotension (one). Hirsutism was troublesome in the two patients remaining on treatment. They concluded that diazoxide may be useful in the management of some patients with primary pulmonary hypertension, but its use is limited by the frequency of side effects.

Several experimental formulations of diazoxide have been tested in humans and animals. These include an oral solution tested in pharmacodynamic and pharmacokinetic studies and a tablet formulation under development in the early 1960's as an anti-hypertensive, but never commercialized (see Calesnick et al., J. Pharm. Sci. 54:1277-1280 (1965); Reddy et al., AAPS Pharm Sci Tech 4(4):1-98, 9 (2003); U.S. Pat. No. 6,361,795).

Current oral formulations of diazoxide are labeled for dosing two or three times per day at 8 or 12 hour intervals. Most subjects receiving diazoxide are dosed three times per day. Commercial and experimental formulations of diazoxide are characterized by rapid drug release following ingestion with complete release in approximately 2 hours. Unless indicated differently, the term “approximately” when used in the context of a numeric value, refer to the stated numeric value +/−10%. In the context of two-theta angles from XRPD studies, the term approximately refers to +/−5% of the stated numeric value.

Current oral formulations of diazoxide in therapeutic use result in a range of adverse side effects including dyspepsia, nausea, diarrhea, fluid retention, edema, reduced rates of excretion of sodium, chloride, and uric acid, hyperglycemia, vomiting, abdominal pain, ileus, tachycardia, palpitations, and headache. (See e.g., current packaging insert for Proglycem®). Oral treatment with diazoxide is used in individuals experiencing serious disease where failure to treat results in significant morbidity and mortality. The adverse side effects from oral administration are tolerated because the benefits of treatment are substantial. The adverse side effects profile of oral diazoxide limit the utility of the drug in treating obese subjects at doses within the labeled range of 3 to 8 mg/kg per day.

The effect of diazoxide in animal models of diabetes and obesity (e.g. obese and lean Zucker rats) has been previously reported. See e.g. Alemzadeh et al., Endocrinology 133:705-712 (1993); Alemzadeh et al., Metabolism 45:334-341 (1996); Alemzadeh et al., Endocrinology 140:3197-3202 (1999); Stanridge et al., FASEB J 14:455-460 (2000); Alemzadeh et al., Med Sci Monit 10(3): BR53-60 (2004); Alemzadeh et al., Endocrinology 145(12):3476-3484 (2004); Aizawa et al., J of Pharma Exp Ther 275(1): 194-199 (1995); and Surwit et al., Endocrinology 141:3630-3637 (2000).

The effect of diazoxide in humans with obesity or diabetes has been previously reported. See e.g., Wigand et al., Diabetes 28(4):287-291 (1979), evaluation of diazoxide on insulin receptors; Ratzmann et al., Int J Obesity 7(5):453-458 (1983), glucose tolerance and insulin sensitivity in moderately obese patients; Marugo et al., Boll Spec It Biol Sper 53:1860-1866 (1977), moderate dose diazoxide treatment on weight loss in obese patients; Alemzadeh et al., J Clin Endocr Metab 83:1911-1915 (1998), low dose diazoxide treatment on weight loss in obese hyperinsulinemic patients; Guldstrand et al., Diabetes and Metabolism 28:448-456 (2002), diazoxide in obese type II diabetic patients; Ortqvist et al., Diabetes Care 27(9):2191-2197 (2004), beta-cell function measured by circulating C-peptide in children at clinical onset of type 1 diabetes; Bjork et al., Diabetes Care 21(3):427-430 (1998), effect of diazoxide on residual insulin secretion in adult type I diabetes patients; and Qvigstad et al., Diabetic Medicine 21:73-76 (2004).

The effect of potassium channel openers on lipid levels has been reported. Gutman et al. (Horm Metab Res 1985 17(10):491-494) studied the effect of diazoxide on an animal model with elevated triglycerides. Normal Wistar rats fed an isocaloric, sucrose-rich (63%) diet (SRD), were reported to develop glucose intolerance and elevated triglyceride levels in plasma (P) as well as in heart (H) and liver (L) tissue. This metabolic state was reported to be accompanied by hyperinsulinism both in vivo and in vitro, consistent with a state of insulin resistance. Gutman et al., administered diazoxide (120 mg/kg/day) together with the diet (SRD+DZX) for 22 days. Control groups fed a standard chow (STD) or the STD plus diazoxide (STD+DZX) were included in the study. Gutman et al., suggested that diazoxide could prevent the development of hyperinsulinism, glucose intolerance and elevated levels of triacylglycerol in plasma, heart and liver present in animals fed on a sucrose rich diet.

Yokoyama et al. (Gen Pharmacol 1998 30(2):233-237) studied the effects of KRN4884 on lipid metabolism in hyperlipidemic rats. KRN4884 is a novel pyridinecarboxamidine type potassium channel opener. Oral administration of KRN4884 (1-10 mg/kg/day) for 14 days was reported to dose dependently reduce serum triglyceride levels in Zucker rats. The reductions in serum triglyceride were associated with reductions in triglyceride in chylomicron and very low density lipoprotein. KRN4884 produced no change in serum insulin and glucose levels in Zucker rats. KRN4884 exhibited a similar triglyceride lowering effect in diet-induced hyperlipidemic rats. In a second study with KRN4884 (J Cardiovasc Pharmacol 2000 35(2):287-293), these authors used high-fructose diet rats which developed hypertension, hypertriglyceridemia, increased total cholesterol/HDL (high-density lipoprotein)-cholesterol ratio, and hyperinsulinemia, and reported that KRN4884 treatment significantly increased lipoprotein lipase (LPL) activity in muscle and tended to increase LPL activity in adipose tissue. Hepatic triglyceride lipase activity was not affected by KRN4884 administration.

Matzno et al. (J Pharmacol Exp Ther 1994 271(3):1666-71) studied the effect of (+)-N-(6-amino-3-pyridil)-N′-[(1S,2R,4R)-bicyclo-[2.2.1]hept-2-yl]-N″-cyanoguanidine hydrochloride (AL0671), a cyanoguanidine-derivative potassium channel opener, on serum lipid and lipoprotein levels in obese Zucker rats. Serial administration (for 1 or 2 weeks) of AL0671 (5 mg/kg/day) was reported to significantly decrease serum total triglyceride, chylomicron and very-low-density lipoprotein levels with increasing high-density lipoprotein cholesterol, whereas low-density lipoprotein levels did not change. AL0671 (5 mg/kg/day) also was reported to increase lipoprotein lipase activities 4-fold and hepatic triglyceride lipase activities 3-fold in postheparin plasma. The authors suggested that AL0671 activates both lipoprotein lipase and hepatic triglyceride lipase activities through its potassium channel-opening activity followed by decreasing triglyceride-rich lipoproteins in genetically obese hyperlipemic rats.

Alemzadeh and Tushaus (Med Sci Monit, 2005; 11(12): BR439-448) studied the effect of 8 weeks of diazoxide treatment (150 mg/kg/day) on triglyceride biosynthesis in Zucker Diabetic Fatty (ZDF) rats. They reported that diazoxide treatment significantly reduced expression of sterol regulatory element-binding protein-1c, fatty acid synthase, acetyl CoA carboxylase, hormone-sensitive lipase, and peroxisome proliferator agonist receptor-γ, without altering expressions of acyl CoA oxidase, peroxisome proliferator receptor-α, and carnitine palmitoyl transferase-1. Also reported was that diazoxide treatment decreased hepatic triglycerides, long chain acyl-CoA and cholesterol contents.

U.S. Pat. No. 5,284,845 describes a method for normalizing blood glucose and insulin levels in an individual exhibiting normal fasting blood glucose and insulin levels and exhibiting in an oral glucose tolerance test, elevated glucose levels and at least one insulin level abnormality selected from the group consisting of a delayed insulin peak, an exaggerated insulin peak and a secondary elevated insulin peak. According to this reference, the method includes administering diazoxide in an amount from about 0.4 to about 0.8 mg/kg body weight before each meal in an amount effective to normalize the blood glucose and insulin levels.

U.S. Pat. No. 6,197,765 describes administration of diazoxide for treatment for syndrome-X, and resulting complications, that include hyperlipidemia, hypertension, central obesity, hyperinsulinemia and impaired glucose tolerance. According to this reference, diazoxide interferes with pancreatic islet function by ablating endogenous insulin secretion resulting in a state of insulin deficiency and high blood glucose levels equivalent to that of diabetic patients that depend on exogenous insulin administration for normalization of their blood glucose levels.

U.S. Pat. No. 2,986,573 describes the preparation of diazoxide and its use for the treatment of hypertension. The patent asserts that alkali metal salts may be prepared by methods well-known in the art for the preparation of a salt of a strong base with a weak acid. It also alleges a specific method for making a sodium salt of diazoxide. This patent does not provide any evidence to support the formation of any salt of diazoxide.

U.S. Pat. No. 5,629,045 describes diazoxide for topical ophthalmic administration.

WO 98/10786 describes use of diazoxide in the treatment of X-syndrome including obesity associated therewith.

U.S. Patent publication no. 2003/0035106 describes diazoxide containing compounds for reducing the consumption of fat-containing foods.

U.S. Patent Publication No. 2004/0204472 describes the use of a Cox-2 inhibitor plus diazoxide in the treatment of obesity. Also described therein is the use of a Cox-2 inhibitor plus a pharmaceutically acceptable salt of diazoxide, wherein acceptable cations include alkali metals and alkaline earth metals.

U.S. Patent Publication No. 2002/0035106 describes use of K_(ATP) channel agonists for reducing the consumption of fat containing food. This application mentions pharmaceutically acceptable acid addition salts, pharmaceutically acceptable metal salts and optionally alkylated ammonium salts, but does not disclose or describe how to prepare any such salts. This patent also does not provide any evidence to support the formation of any salt of a K_(ATP) channel agonist.

U.K. Patent GB982072 describes the preparation and use of diazoxide and derivatives for the treatment of hypertension and peripheral vascular disorders. This patent mentions non-toxic alkali metals salts but does not disclose or describe how to prepare any such salts. This patent does not provide any evidence to support the formation of any salt of diazoxide or its derivatives.

SUMMARY OF THE INVENTION

The current invention relates to methods of preparation and use of formulations that include alkali metal, tertiary amine and ammonium salts of diazoxide and diazoxide derivatives. It has been surprisingly found that it is difficult to produce salts of diazoxide and derivatives. In particular, the inventors have been unable to reproduce formation of a diazoxide salt using the method asserted in U.S. Pat. No. 2,986,573. Contrary to what is reported in the literature, salt formation with diazoxide and derivatives depends on a proper selection of solvent and counter-ion.

Provided herein are pharmaceutical formulations of K_(ATP) channel openers and their use for treatment of various diseases and conditions including but not limited to type 1 and type 2 diabetes, hypertension, dyslipidemia, nonalcoholic steatohepatitis (NASH) pulmonary hypertension, myocardial infarction and arrhythmias following myocardical infarction, and poly-cystic ovarian syndrome. Such formulations are characterized as being bioavailable. A K_(ATP) channel opener as used herein has any one or more of the following properties: (1) opening SURx/Kir6.y potassium channels, where x=1, 2A or 2B and y=1 or 2; (2) binding to the SURx subunit of K_(ATP) channels; and (3) inhibiting glucose induced release of insulin following administration of the compound in vivo. Preferably, K_(ATP) channel openers are K_(ATP) channel openers with all three properties. K_(ATP) channel openers as defined herein are preferably salts prepared from the compounds of Formulae I-VIII, as set forth below.

In another aspect, the present invention also provides salts of the compounds defined by Formulae I-VIII. Salts of Formulae I-IV provided herein include monovalent alkali metal salts and monovalent and divalent salts of organic compounds, preferably organic compounds which include an ammonium moiety. Salts of Formulae V-VIII are also provided herein, preferably prepared with monovalent and divalent counter-ions.

K_(ATP) channel openers defined by Formula I are as follows:

wherein:

-   -   R¹ is selected from the group consisting of hydrogen, lower         alkyl, substituted lower alkyl, cycloalkyl, and substituted         cycloalkyl provided however that when R¹ is a substituted lower         alkyl or a substituted cycloalkyl, then the substituent does not         include an amino group;     -   R^(2a) is hydrogen;     -   X is a 1, 2 or 3 atom chain, wherein each atom is independently         selected from carbon, sulfur and nitrogen, and each atom is         optionally substituted with halogen, hydroxyl, lower alkyl,         substituted lower alkyl, lower alkoxy, cycloalkyl, substituted         cycloalkyl, or substituted lower alkoxy, provided however that         when an atom of the chain is substituted with substituted lower         alkyl, substituted lower alkoxy or substituted cycloalkyl, then         the substituent does not include an amino group;     -   wherein ring B is saturated, monounsaturated, polyunsaturated or         aromatic;     -   and all bioequivalents including salts, prodrugs and isomers         thereof.

In particular embodiments of Formula I, X is C(R^(a))C(R^(b)), wherein R^(a) and R^(b) are independently selected from the group consisting of hydrogen, halogen, lower alkyl, substituted lower alkyl, cycloalkyl, substituted cycloalkyl, lower alkoxy, substituted lower alkoxy, sulfonyl, and the like. In further embodiments, R^(a) and R^(b) are independently selected from the group consisting of hydroxyl, substituted oxy, substituted thiol, alkylthio, substituted alkylthio, sulfinyl, sulfonyl, substituted sulfinyl, substituted sulfonylalkylsulfinyl, alkylsulfonyl, and the like. In a preferred embodiment, Ring B does not include any heteroatoms.

Salts of embodiments of the channel openers defined by Formula I may be prepared from the following: (a) metal hydroxides, preferably alkali metal hydroxides (e.g., NaOH and KOH) and (b) organic hydroxides, preferably organic compounds which include at least one tertiary amine or at least one quaternary ammonium ion (e.g., diethylaminoethanol, triethylamine, hydroxyethylpyrrolidine, choline and hexamethylhexamethylenediammonium, and the like).

K_(ATP) channel openers defined by Formula II are as follows:

wherein:

-   -   R¹ is selected from the group consisting of hydrogen, lower         alkyl, substituted lower alkyl, cycloalkyl, and substituted         cycloalkyl provided however that when R¹ is a substituted lower         alkyl or a substituted cycloalkyl, then the substituent does not         include an amino group;     -   R^(2b) is hydrogen;     -   X is a 1, 2 or 3 atom chain, wherein each atom is independently         selected from carbon, sulfur and nitrogen, and each atom is         optionally substituted with halogen, hydroxyl, lower alkyl,         substituted lower alkyl, lower alkoxy, cycloalkyl, substituted         cycloalkyl, or substituted lower alkoxy, provided however that         when an atom of the chain is substituted with substituted lower         alkyl, substituted cycloalkyl or substituted lower alkoxy, then         the substituent does not include an amino group;     -   wherein ring B is saturated, monounsaturated, polyunsaturated or         aromatic;     -   and all bioequivalents including salts, prodrugs and isomers         thereof.

In particular embodiments of Formula II, X is C(R^(a))C(R^(b)), wherein R^(a) and R^(b) are independently selected from the group consisting of hydrogen, halogen, lower alkyl, substituted lower alkyl, cycloalkyl, substituted cycloalkyl, lower alkoxy, substituted lower alkoxy, sulfonyl, and the like. In further embodiments, R^(a) and R^(b) are independently selected from the group consisting of hydroxyl, substituted oxy, substituted thiol, alkylthio, substituted alkylthio, sulfinyl, sulfonyl, substituted sulfinyl, substituted sulfonyl, alkylsulfinyl, alkylsulfonyl, nitro and the like. In preferred embodiment, Ring B does not include any heteroatoms.

Salts of embodiments of the channel openers defined by Formula II may be prepared from the following: (a) metal hydroxides, preferably alkali metal hydroxides (e.g., NaOH and KOH) and (b) organic hydroxides, preferably organic compounds which include at least one tertiary amine or at least one quaternary ammonium ion (e.g., diethylaminoethanol, triethylamine, hydroxyethylpyrrolidine, choline and hexamethylhexamethylenediammonium, and the like).

K_(ATP) channel openers defined by Formula III are as follows:

wherein:

-   -   R¹ is selected from the group consisting of hydrogen, lower         alkyl, substituted lower alkyl, and cycloalkyl provided however         that when R¹ is a substituted lower alkyl, then the substituent         does not include an amino group;     -   R^(2a) is hydrogen;     -   R³ is selected from the group consisting of hydrogen, halogen,         lower alkyl, substituted lower alkyl, cycloalkyl and substituted         cycloalkyl provided however that when R³ is a substituted lower         alkyl, then the substituent does not include an amino group;     -   R⁴ is selected from the group consisting of hydrogen, halogen,         lower alkyl, substituted lower alkyl, cycloalkyl and substituted         cycloalkyl provided however that when R⁴ is a substituted lower         alkyl, then the substituent does not include an amino group;     -   and all bioequivalents including salts, prodrugs and isomers         thereof.

In particular embodiments of Formula III, R¹ is a lower alkyl, (preferably ethyl or methyl); R^(2a) is hydrogen; and R³ and R⁴ are each independently halogen.

In another embodiment of Formula III, R¹ is methyl; R^(2a) is hydrogen; R³ is selected from the group consisting of hydrogen, halogen, lower alkyl, substituted lower alkyl, cycloalkyl, and substituted cycloalkyl; and R⁴ is chlorine.

Salts of embodiments of the channel openers defined by Formula III may be prepared from the following: (a) metal hydroxides, preferably alkali metal hydroxides (e.g., NaOH and KOH) and (b) organic hydroxides, preferably organic compounds which include at least one tertiary amine or at least one quaternary ammonium ion (e.g., diethylaminoethanol, triethylamine, hydroxyethylpyrrolidine, choline and hexamethylhexamethylenediammonium, and the like).

K_(ATP) channel openers defined by Formula IV are as follows:

wherein:

-   -   R¹ is selected from the group consisting of hydrogen, lower         alkyl, substituted lower alkyl, and cycloalkyl provided however         that when R¹ is a substituted lower alkyl, then the substituent         does not include an amino group;     -   R^(2b) is hydrogen;     -   R³ is selected from the group consisting of hydrogen, halogen,         lower alkyl, substituted lower alkyl, cycloalkyl and substituted         cycloalkyl provided however that when R³ is a substituted lower         alkyl, then the substituent does not include an amino group;     -   R⁴ is selected from the group consisting of hydrogen, halogen,         lower alkyl, substituted lower alkyl, cycloalkyl and substituted         cycloalkyl provided however that when R⁴ is a substituted lower         alkyl, then the substituent does not include an amino group;     -   and all bioequivalents including salts, prodrugs and isomers         thereof.

In particular embodiments of Formula IV, R¹ is a lower alkyl, (preferably ethyl or methyl); R^(2b) is hydrogen; and R³ and R⁴ are each independently halogen.

In another embodiment of Formula IV, R¹ is methyl; R^(2b) is hydrogen; R³ is selected from the group consisting of hydrogen, halogen, lower alkyl, substituted lower alkyl, cycloalkyl, and substituted cycloalkyl; and R⁴ is chlorine.

Salts of embodiments of the channel openers defined by Formula IV may be prepared from the following: (a) metal hydroxides, preferably alkali metal hydroxides (e.g., NaOH and KOH) and (b) organic hydroxides, preferably organic compounds which include at least one tertiary amine or at least one quaternary ammonium ion (e.g., diethylaminoethanol, triethylamine, hydroxyethylpyrrolidine, choline and hexamethylhexamethylenediammonium, and the like).

K_(ATP) channel openers defined by Formula V are as follows:

wherein:

-   -   R¹ is selected from the group consisting of optionally         substituted amino, optionally substituted alkyl, optionally         substituted cycloalkyl, optionally substituted heterocyclyl,         optionally substituted heterocyclylalkyl, optionally substituted         aryl, optionally substituted heteroaryl, and optionally         substituted heteroarylalkyl;     -   R^(2a) is selected from the group consisting of hydrogen, and         lower alkyl;     -   X is a 1, 2 or 3 atom chain, wherein each atom is independently         selected from carbon, sulfur and nitrogen, and each atom is         optionally substituted with halogen, hydroxyl, optionally         substituted lower alkyl, optionally substituted lower alkoxy,         optionally substituted cycloalkyl, or optionally substituted         amino;     -   wherein ring B is saturated, monounsaturated, polyunsaturated or         aromatic;     -   wherein at least one of R¹ or a substituent of X includes an         amino group;     -   and all bioequivalents including salts, prodrugs and isomers         thereof.

In particular embodiments of Formula V, X is C(R^(a))C(R^(b)), wherein R^(a) and R^(b) are independently selected from the group consisting of hydrogen, halogen, optionally substituted lower alkyl, optionally substituted cycloalkyl, optionally substituted lower alkoxy, amino, sulfonylamino, aminosulfonyl, sulfonyl, and the like. Preferably R¹ includes at least one substituent containing an amino group. In further embodiments, R^(a) and R^(b) are independently selected from the group consisting of hydroxyl, substituted oxy, substituted thiol, alkylthio, substituted alkylthio, sulfinyl, sulfonyl, substituted sulfinyl, substituted sulfonyl, substituted sulfonylamino, substituted amino, substituted amine, alkylsulfinyl, alkylsulfonyl, alkylsulfonylamino, and the like. In a preferred embodiment, Ring B does not include any heteroatoms.

K_(ATP) channel openers defined by Formula VI are as follows:

wherein:

-   -   R¹ is selected from the group consisting of optionally         substituted amino, optionally substituted alkyl, optionally         substituted cycloalkyl, optionally substituted heterocyclyl,         optionally substituted heterocyclylalkyl, optionally substituted         aryl, optionally substituted heteroaryl, and optionally         substituted heteroarylalkyl;     -   R^(2b) is selected from the group consisting of hydrogen and         lower alkyl;     -   X is a 1, 2 or 3 atom chain, wherein each atom is independently         selected from carbon, sulfur and nitrogen, and each atom is         optionally substituted with halogen, hydroxyl, optionally         substituted lower alkyl, optionally substituted lower alkoxy,         optionally substituted cycloalkyl, or optionally substituted         amino;     -   wherein ring B is saturated, monounsaturated, polyunsaturated or         aromatic;     -   wherein at least one of R¹ or a substituent of X includes an         amino group;     -   and all bioequivalents including salts, prodrugs and isomers         thereof.

In particular embodiments of Formula VI, X is C(R^(a))C(R^(b)), wherein R^(a) and R^(b) are independently selected from the group consisting of hydrogen, halogen, lower alkyl, substituted lower alkyl, cycloalkyl, substituted cycloalkyl, lower alkoxy, substituted lower alkoxy, amino, sulfonylamino, aminosulfonyl, sulfonyl, and the like. In further embodiments, R^(a) and R^(b) are independently selected from the group consisting of hydroxyl, substituted oxy, substituted thiol, alkylthio, substituted alkylthio, sulfinyl, sulfonyl, substituted sulfinyl, substituted sulfonyl, substituted sulfonylamino, substituted amino, substituted amine, alkylsulfinyl, alkylsulfonyl, alkylsulfonylamino, and the like. Preferably R¹ includes at least one substituent containing an amino group. In a preferred embodiment, Ring B does not include any heteroatoms.

K_(ATP) channel openers defined by Formula VII are as follows:

wherein:

-   -   R¹ is selected from the group consisting of optionally         substituted amino, optionally substituted alkyl, optionally         substituted cycloalkyl, optionally substituted heterocyclyl,         optionally substituted heterocyclylalkyl, optionally substituted         aryl, optionally substituted heteroaryl, and optionally         substituted heteroarylalkyl;     -   R^(2a) is selected from the group consisting of hydrogen, lower         alkyl, and substituted lower alkyl;     -   R³ is selected from the group consisting of hydrogen, halogen,         optionally substituted lower alkyl, optionally substituted         amino, optionally substituted cycloalkyl and optionally         substituted aryl;     -   R⁴ is selected from the group consisting of hydrogen, halogen,         optionally substituted lower alkyl, optionally substituted         amino, optionally substituted cycloalkyl and optionally         substituted aryl;     -   wherein at least one of R¹, R³ and R⁴ includes a substituent         containing an amino group;     -   and all bioequivalents including salts, prodrugs and isomers         thereof.

Preferably, R¹ includes a substituent containing an amino group. In particular embodiments of Formula VII; R¹ includes an amino substituent, R^(2a) is hydrogen; and R³ and R⁴ are each independently halogen.

In another embodiment of Formula VII, R^(2a) is hydrogen; R³ is selected from the group consisting of hydrogen, halogen, lower alkyl, substituted lower alkyl, amino, substituted amino, cycloalkyl, and substituted cycloalkyl; and R⁴ is chlorine.

K_(ATP) channel openers defined by Formula VIII are as follows:

wherein:

-   -   R¹ is selected from the group consisting of optionally         substituted amino, optionally substituted alkyl, optionally         substituted cycloalkyl, optionally substituted heterocyclyl,         optionally substituted heterocyclylalkyl, optionally substituted         aryl, optionally substituted heteroaryl, and optionally         substituted heteroarylalkyl;     -   R^(2b) is selected from the group consisting of hydrogen, lower         alkyl, and substituted lower alkyl;     -   R³ is selected from the group consisting of hydrogen, halogen,         optionally substituted lower alkyl, optionally substituted         amino, optionally substituted cycloalkyl and optionally         substituted aryl;     -   R⁴ is selected from the group consisting of hydrogen, halogen,         optionally substituted lower alkyl, optionally substituted         amino, optionally substituted cycloalkyl and optionally         substituted aryl;     -   wherein at least one of R¹, R³ and R⁴ includes a substituent         containing an amino group;     -   and all bioequivalents including salts, prodrugs and isomers         thereof.

Preferably R¹ includes a substituent containing an amino group. In particular embodiments of Formula VII, R^(2b) is hydrogen; and R³ and R⁴ are each independently halogen.

In another embodiment of Formula VII, R^(2b) is hydrogen; R³ is selected from the group consisting of hydrogen, halogen, lower alkyl, optionally substituted lower alkyl, optionally substituted amino, and optionally substituted cycloalkyl; and R⁴ is chlorine.

Unless otherwise indicated, reference in this application to K_(ATP) channel openers should be understood to refer to K_(ATP) channel openers based upon a salt of one of the compounds described by Formulae I-VIII and having one or more, and preferably all three, of the following properties: (1) opening SURx/Kir6.y potassium channels, wherein x=1, 2A or 2B and y=1 or 2; (2) binding to the SURx subunit of K_(ATP) channels; and (3) inhibiting glucose induced release of insulin following administration of the compound in vivo. Such K_(ATP) channel openers preferably have the structure of any of the compounds of Formula I-VIII, or more preferably Formula III-IV where ring B or its equivalent does not include any heteroatoms. More preferably, the structure is diazoxide. Structural variants or bioequivalents of any of the compounds defined by Formulae I-VIII, such as derivatives, salts, prodrugs or isomers, are also contemplated herein. Specifically, salts of compounds of Formula I-IV wherein the cation is selected from a cation of an alkali metal or an organic compound which includes a tertiary amine or a quaternary ammonium ion. Preferably, when the salt includes an anion of diazoxide and a sodium cation, the salt is not in a form suitable for intravenous use. In other embodiments, when the anion is diazoxide in a solution suitable for intravenous use, the cation is not sodium. In alternate embodiments, in solutions suitable for intravenous use, when the cation is sodium, the anion is not an anion of diazoxide. In certain embodiments, when the salt includes an anion of diazoxide and a sodium cation, the salt is not in liquid form. More preferably, K_(ATP) channel openers contemplated herein are salts of compounds of Formulae III and IV wherein the cation is selected from sodium, potassium, choline or hexamethyl hexamethylene diammonium. Other K_(ATP) channel openers that are contemplated for use herein include BPDZ 62, BPDZ 73, NN414, BPDZ 154.

Also provided herein are salts of compounds of Formula V-VIII, wherein at least one substituent of the compound of Formulae V-VIII includes an amino group. In another embodiment, the compound of Formula V-VIII forms the anion of the salt and a monovalent or divalent metal forms the cation. In other embodiments, the cation includes a tertiary amino or quaternary ammonium group.

In vitro analysis of glucose induced release of insulin via K_(ATP) channel openers can be determined using rat islets as provided by De Tullio et al., J. Med. Chem., 46:3342-3353 (2003), or by using human islets as provided by Björklund et al., Diabetes, 49:1840-1848 (2000).

Provided herein are formulations, such as controlled release pharmaceutical formulations, of K_(ATP) channel openers and bioequivalents thereof, which include salts of the compounds of Formulae I-VIII. In one embodiment, the salt can be formulated for controlled release following oral administration. Such formulations contain in a single administration dosage between 10 and 100 mg, between 25 and 100 mg, between 100 and 200 mg, between 200 and 300 mg, between 300 and 500 mg or between 500 and 2000 mg of the salt of the K_(ATP) channel openers provided in Formulae I-VIII. In certain embodiments, the dosage of the K_(ATP) channel openers contained in a formulation may be determined based on the weight of the subject for which it is to be administered, i.e., the formulation may contain in a single administration dosage between 0.1-20 mg of the K_(ATP) channel opener per kg of the subject's body weight, or between 0.1-0.5 mg of the K_(ATP) channel opener per kg of the subject's body weight; or between 0.5-1 mg of the K_(ATP) channel opener per kg of the subject's body weight; or between 1-2 mg of the K_(ATP) channel opener per kg of the subject's body weight, or between 2-5 mg of the K_(ATP) channel opener per kg of the subject's body weight, or between 5-10 mg of the K_(ATP) channel opener per kg of the subject's body weight, or between 10-15 mg of the K_(ATP) channel opener per kg of the subject's body weight, or between 15-20 mg of the K_(ATP) channel opener per kg of the subject's body weight.

Also provided herein are controlled release pharmaceutical formulations containing K_(ATP) channel openers selected from salts of Formulae I-VIII, which can be obtained by at least one of the following: (a) particle size reduction involving comminution, spray drying, or other micronising techniques, (b) use of an ion exchange resin, (c) use of inclusion complexes, for example cyclodextrin, (d) compaction of the K_(ATP) channel opener with a solubilizing agent including a low viscosity hypromellose, low viscosity methylcellulose or similarly functioning excipient or combinations thereof, (e) associating the K_(ATP) channel opener with a salt prior to formulation, (f) use of a solid dispersion of the K_(ATP) channel opener, (g) use of a self emulsifying system, (h) addition of one or more surfactants to the formulation, (i) use of nanoparticles, or (j) combinations of these approaches.

Further provided herein are controlled release pharmaceutical formulations containing K_(ATP) channel openers selected from salts of the compounds defined by Formulae I-VIII, which include at least one component that substantially inhibits release of the K_(ATP) channel activator from the formulation until after gastric transit. As used herein, “substantially inhibits” means less than 15% release, more preferably at least less than 10% release, or even more preferably at least less than 5% release of the drug from the formulation during gastric transport. Release can be measured in a standard USP based in-vitro gastric dissolution assay in a calibrated dissolution apparatus. See e.g., U.S. Pharmacopeia, Chapter 711 (2005).

Also provided are oral pharmaceutical formulations of the K_(ATP) channel openers selected from the salts of the compounds of Formulae I-VIII, which include at least one component that substantially inhibits release of the K_(ATP) channel opener from the formulation until after gastric transit. Substantial inhibition of drug release during gastric transit is achieved by inclusion of a component in the formulation selected from the group consisting of: (a) a pH sensitive polymer or co-polymer applied as a compression coating on a tablet, (b) a pH sensitive polymer or co-polymer applied as a thin film on a tablet, (c) a pH sensitive polymer or co-polymer applied as a thin film to an encapsulation system, (d) a pH sensitive polymer or co-polymer applied to encapsulated microparticles, (e) a non-aqueous-soluble polymer or copolymer applied as a compression coating on a tablet, (f) a non-aqueous-soluble polymer or co-polymer applied as a thin film on a tablet, (g) a non-aqueous soluble polymer applied as a thin film to an encapsulation system, and (h) a non-aqueous soluble polymer applied to microparticles, wherein the pH sensitive polymer or co-polymer is resistant to degradation under acid conditions. Alternatively, substantial inhibition of drug release during gastric transport can also be achieved by incorporation of the formulation in an osmotic pump system, by use of systems controlled by ion exchange resins, or by combinations of any of the above approaches.

Also provided herein are controlled release pharmaceutical formulations of K_(ATP) channel openers selected from salts of the compounds of Formulae I-VIII, wherein the formulation includes at least one component that contributes to sustained release of a K_(ATP) channel opener over an extended period, e.g., over a period of 2-24 hours following administration, or over a period of 2-4 hours following administration, or over a period of 4-8 hours following administration, or over a period of more than 8-24 hours following administration. These formulations are characterized in having one of the following components: (a) a pH sensitive polymeric coating, (b) a hydrogel coating, (c) a film coating that controls the rate of diffusion of the drug from a coated matrix, (d) an erodable matrix that controls rate of drug release, (e) polymer coated pellets, granules or microparticles of drug which can be further encapsulated or compressed into a tablet, (f) an osmotic pump system containing the drug, (g) a compression coated tablet form of the drug, or (h) combinations of any of the approaches of (a)-(f) above.

As used herein, an erodable matrix is the core of a tablet formulation that, upon exposure to a suitable aqueous environment, begins a process of disintegration which facilitates the release of drug from the matrix. The rate of release of drug from the tablet is controlled both by the solubility of the drug and the rate of disintegration of the matrix.

The above formulations may further comprise one or more additional pharmaceutically active agents (other than K_(ATP) channel openers selected from the salts of the compounds of Formulae I-VIII) useful for the treatment of a condition selected from the group consisting of obesity, prediabetes, diabetes, hypertension, depression, elevated cholesterol, fluid retention, other obesity associated co-morbidities, ischemic and reperfusion injury, epilepsy, cognitive impairment, schizophrenia, mania, other psychotic diseases, and the like.

Further provided is a controlled release pharmaceutical formulation of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII wherein administration to an obese, overweight or obesity prone subject results in at least one of the following: (a) inhibition of fasting insulin secretion, (b) inhibition of stimulated insulin secretion, (c) elevation of energy expenditure, (d) elevation of beta oxidation of fat, or (e) inhibition of hyperphagia for about 24 hours.

Additionally provided is a controlled release pharmaceutical formulation of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII wherein administration to an obese, overweight or obesity prone subject results in at least one of the following: (a) inhibition of fasting insulin secretion, (b) inhibition of glucose stimulated insulin secretion, (c) elevation of energy expenditure, (d) elevation of beta oxidation of fat, or (e) inhibition of hyperphagia for about 18 hours.

Still further provided is a controlled release pharmaceutical formulation of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII which upon administration to an obese, overweight or obesity prone subject results in at least one of the following: (a) inhibition of fasting insulin secretion, (b) inhibition of glucose stimulated insulin secretion, (c) elevation of energy expenditure, (d) elevation of beta oxidation of fat, or (e) inhibition of hyperphagia for about 24 hours.

Additionally provided is a controlled release pharmaceutical formulation of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII that upon administration to an obese, overweight or obesity prone subject results in at least one of the following: (a) inhibition of fasting insulin secretion, (b) inhibition of glucose stimulated insulin secretion, (c) elevation of energy expenditure, (d) elevation of beta oxidation of fat, or (e) inhibition of hyperphagia for about 18 hours.

In other embodiments of the invention there are provided formulation comprising diazoxide choline and about 1% to about 55% by weight of a polymer as described herein. For example, the polymer may be selected from the group consisting of polyethylene oxide and cellulose. Cellulose suitable for use in such formulations may be selected from hydroxypropylmethyl cellulose, hydroxypropylcellulose, ethylcellulose, methylcellulose, carboxymethylcellulose, and a mixture of any two or more thereof. Various polyethylene oxides may be used in the formulations, including those selected from PEO N750, PEO 303 or a mixture thereof.

Provided herein are methods of using any of the salts of the compounds of Formulae I-VIII and formulations thereof. For example, provided herein is a method of treating hypoglycemia, the method comprising orally administering to a subject in need thereof, a controlled release formulation of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII.

Further provided herein is a method of treating obesity associated co-morbidities in an obese, overweight or obesity prone subject, the method comprising administering a therapeutically effective amount of a solid oral dosage form of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII, or controlled release pharmaceutical formulation of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII. In a preferred embodiment, administration is no more than two times per 24 hours, or once per 24 hours.

Yet further provided herein is a method of achieving weight loss in an obese overweight, or obesity prone subject, the method comprising administering a therapeutically effective amount of a solid oral dosage form of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII or controlled release pharmaceutical formulation of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII. In a preferred embodiment, administration is no more than two times per 24 hours, or once per 24 hours. The daily dosage administered is preferably between 50 and 180 mg. In certain embodiments, the obese subject has a body mass index greater than 30 kg/m², or greater than 35 kg/m², or greater than 40 kg/m², or greater than 50 g/m², or greater than 60 kg/m² at the time the method commences.

Also provided is a method of maintaining a weight loss in an obese overweight, or obesity prone subject, the method comprising administering a therapeutically effective amount of a solid oral dosage form of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII or controlled release pharmaceutical formulation of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII. It is preferable to maintain weight in an obese subject once some weight loss has occurred when the alternative is to regain weight. In a preferred embodiment, administration is no more than two times per 24 hours, or once per 24 hours.

Also provided is a method of treating obesity or type 1 or type 2 diabetes with a salt of KATP channel opener which minimizes a range of adverse events including but not limited to headaches, fluid retention, edema, hyperglycemia, and tachycardia. In accordance with this method, the initial dose is chosen to provide for elevations in supine heart rate that on average are not greater than 5 beats per minute, followed by tolerization and a return to baseline heart rate by the time steady state circulating K_(ATP) channel opener levels are reached. This starting dose is preferably between 10 and 200 mg of K_(ATP) channel opener or the equivalent based on relative bioavailability and potency compared to diazoxide. Preferably, the K_(ATP) channel opener is diazoxide. The starting dose is given to the patient daily for days or a few weeks. In further embodiments, the method includes escalating the dose which is administered when after supine heart rates return to baseline. An initial escalating dose is established using the same criteria as the initial dose. The initial escalating dose is given to the patient daily for days or a few weeks. The increment in dose is preferentially 25 to 150 mg of K_(ATP) channel opener or the equivalent based on relative bioavailability and potency compared to diazoxide. Further rounds of this dose escalation would continue until the therapeutically effective dose is reached. Alternatively, standing heart rate may be utilized where the average increase is not greater than 10 beats per minute.

Further provided is a method of treating subjects with type 1 or type 2 diabetes suffering from hypoglycemia associated autonomic failure involving the administration of an agent that either stimulates glucagon release from pancreatic alpha cells via interaction with K_(ATP) channels and/or stimulates hypothalamic neurons resulting in an amplification of or restoration of the counter regulatory hormonal response to hypoglycemia, via activation of K_(ATP) channels. In a preferred embodiment, the agent is a therapeutically effective amount of a formulation selected from the group consisting of:

i) a formulation comprising a salt, wherein the salt comprises an anion of a K_(ATP) channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising a tertiary amine or ammonium group;

ii) a formulation comprising a salt, wherein the salt comprises an anion of a K_(ATP) channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII; and

iii) a formulation comprising a salt, wherein the salt comprises an anion of a K_(ATP) channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII, wherein at least one substituent of the compound of Formulae V-VIII includes an amino group. In another embodiment, the compound of Formula V-VIII forms the anion of the salt and a monovalent or divalent metal forms the cation. In other embodiments, the cation includes a tertiary amino or quaternary ammonium group.

Further provided is a method of elevating energy expenditure in an overweight, obese or obesity prone subject, the method comprising administering an effective amount of a solid oral dosage form of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII or controlled release pharmaceutical formulation of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII. In a preferred embodiment, administration is no more than two times per 24 hours, or once per 24 hours. In certain embodiments, the subject has a body mass index greater than 20 kg/m², or greater than 25 kg/m², or greater than 30 kg/m², or greater than 35 kg/m², or greater than 40 kg/m², or greater than 50 kg/m², or greater than 60 kg/m² at the time the method commences.

Additionally provided is a method of elevating beta oxidation of fat in an overweight, obese or obesity prone subject, the method comprising administering an effective amount of a solid oral dosage form of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII or controlled release pharmaceutical formulation of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII. In a preferred embodiment, administration is no more than two times per 24 hours, or once per 24 hours. In certain embodiments, the subject has a body mass index greater than 20 kg/m², or greater than 25 kg/m², or greater than 30 kg/m², or greater than 35 kg/m², or greater than 40 kg/m², or greater than 50 kg/m², or greater than 60 kg/m² at the time the method commences.

Yet further provided is a method of reducing visceral fat in an overweight, obese or obesity prone subject, the method comprising administering an effective amount of a solid oral dosage form of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII or controlled release pharmaceutical formulation of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII. In a preferred embodiment, administration is no more than two times per 24 hours, or once per 24 hours.

Still further provided is a method of delaying or preventing the transition to diabetes of a prediabetic subject comprising administering an effective amount of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII or controlled release pharmaceutical formulation of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII. In a preferred embodiment, administration is no more than two times per 24 hours, or once per 24 hours.

Additionally provided is a method of restoring normal glucose tolerance in a prediabetic subject comprising administering an effective amount of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII or controlled release pharmaceutical formulation of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII. In a preferred embodiment, administration is no more than two times per 24 hours, or once per 24 hours.

Further provided is a method of restoring normal glucose tolerance in a diabetic subject comprising administering an effective amount of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII or controlled release pharmaceutical formulation of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII. In a preferred embodiment, administration is no more than two times per 24 hours, or once per 24 hours.

Still further provided is a method of delaying or preventing progression of diabetes in an subject comprising administering an effective amount of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII or controlled release pharmaceutical formulation of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII. In a preferred embodiment, administration is no more than two times per 24 hours, or once per 24 hours.

Also provided is a method to prevent or treat weight gain, impaired glucose tolerance or dyslipidemia associated with the administration of anti-psychotics to a subject, said method including the co-administration of an effective amount of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII or controlled release pharmaceutical formulation of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII. In a preferred embodiment, administration is no more than two times per 24 hours, or once per 24 hours.

Further provided is a method to treat obesity, or hyperphagia in a Prader-Willi Syndrome patient, a Froelich's Syndrome patient, in a Cohen Syndrome patient, in a Summit Syndrome patient, in an Alstrom Syndrome patient, in a Borjeson Syndrome patient or in a Bardet-Biedl Syndrome patient comprising the administration of an effective amount of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII or controlled release pharmaceutical formulation of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII. In a preferred embodiment, administration is no more than two times per 24 hours, or once per 24 hours.

Still further provided is a method to treat obesity or elevated triglycerides in a patient suffering hyperlipoproteinemia type I, type II, type III or type IV comprising administering an effective amount of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII or controlled release pharmaceutical formulation of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII. In a preferred embodiment, administration is no more than two times per 24 hours, or once per 24 hours.

Also provided is a method of reducing the incidence of adverse effects from administration of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII in the treatment of diseases of a subject achieved by any of the following: (a) use of a dosage form that on administration reduces C_(max) relative to the current Proglycem® oral suspension or capsule products in order to reduce the incidence of adverse side effects that are associated with peak drug levels, (b) use of a dosage form that delays release until gastric transit is complete in order to reduce the incidence of adverse side effects that are associated with the release of drug in the stomach, (c) initiating dosing at subtherapeutic levels and in a stepwise manner increasing dose daily until the therapeutic dose is achieved wherein the number of steps is 2 to 10 to reduce the incidence of adverse side effects that occur transiently at the initiation of treatment, (d) use of the lowest effective dose to achieve the desired therapeutic effect in order to reduce the incidence of adverse side effects that are dose dependent, or (e) optimizing the timing of administration of dose within the day and relative to meals.

Further provided is a method of reducing the incidence of adverse effects from administration of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII without substantially impacting the pharmacokinetic profile of said administered K_(ATP) channel opener, comprising administering to a subject said K_(ATP) channel opener orally in conjunction with a meal which includes solid food. As used herein, in administration of the opener in conjunction with a meal means that the two are ingested within 15 minutes of each other. Accordingly, there is provided a method of reducing the incidence of adverse effects resulting from orally administering a salt of a K_(ATP) channel opener comprising, causing to be orally ingested a formulation selected from the group consisting of:

i) a formulation comprising a salt, said salt comprising an anion of a K_(ATP) channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising a tertiary amine or ammonium group;

ii) a formulation comprising a salt, said salt comprising an anion of a K_(ATP) channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII; and

iii) a formulation comprising a salt, said salt comprising an anion of a K_(ATP) channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII, wherein at least one substituent comprises an amino group; and,

a meal containing one or more solid food items sufficient to reduce the incidence of adverse effects resulting from the orally ingested salt of a K_(ATP) channel opener. In a preferred embodiment, the formulation and the meal are taken within 15 minutes or less of each other.

Further provided is a method of preventing weight gain, dyslipidemia or impaired glucose tolerance in a subject treated with an anti-psychotic drug, the method comprising administering a pharmaceutical formulation of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII.

Yet further provided is a method of treating weight gain, dyslipidemia or impaired glucose tolerance in a subject treated with an anti-psychotic drug, the method comprising administering a pharmaceutical formulation of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII.

Also provided is a method of treating diseases characterized by obesity, hyperphagia, dyslipidemia, or decreased energy expenditure including (a) Prader-Willi Syndrome, (b) Froelich's syndrome, (c) Cohen syndrome, (d) Summit Syndrome, (e) Alstrom Syndrome, (f) Borjesen Syndrome, (g) Bardet-Biedl Syndrome, or (h) hyperlipoproteinemia type I, II, III, and IV comprising administering a pharmaceutical formulation of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII.

Further provided is a pharmaceutical formulation of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII further comprising a pharmaceutically active agent other than the K_(ATP) channel opener. In this formulation, the other pharmaceutically active agent is an agent useful for the treatment of a condition selected from the group consisting of obesity, prediabetes, diabetes, hypertension, depression, elevated cholesterol, fluid retention, or other obesity associated co-morbidities, ischemic and reperfusion injury, epilepsy, cognitive impairment, schizophrenia, mania, and other psychotic condition.

The formulations containing K_(ATP) channel openers selected from the salts of the compounds of Formulae I-VIII described herein provide for improved compliance, efficacy and safety, and for co-formulations with other agents. Included are co-formulations of K_(ATP) channel openers selected from the salts of the compounds of Formulae I-VIII with one or more additional pharmaceutically active agents that have complementary or similar activities or targets. Other pharmaceutically active agents that can be combined with K_(ATP) channel openers selected from the salts of the compounds of Formulae I-VIII to treat obesity or to maintain weight loss in an obesity prone subject include, but are not limited to: sibutramine, orlistat, phentermine, rimonabant, a diuretic, an antiepileptic, or other pharmaceutical active whose therapeutic utility includes weight loss. It is preferable to maintain weight in an obese subject once some weight loss has occurred when the alternative is to regain weight. Other pharmaceutically active agents that may be combined with K_(ATP) channel openers selected from the salts of the compounds of Formulae I-VIII to treat type II diabetes, or prediabetes include acarbose, miglitol, metformin, repaglinide, nateglinide, rosiglitazone, proglitazone, ramipril, metaglidasen, or any other pharmaceutical active that improves insulin sensitivity or glucose utilization or glycemic control where the mode of action is not enhanced insulin secretion. Other pharmaceutical active agent that can be combined with K_(ATP) channel openers selected from the salts of the compounds of Formulae I-VIII to treat obesity associated co-morbidities include a drug active used to lower cholesterol, a drug active used to lower blood pressure, an anti-inflammatory drug that is not a cox-2 inhibitor, a drug that is an antidepressant, a drug used to treat urinary incontinence, or other drug routinely used to treat disease conditions the incidence of which is elevated in overweight or obese patients as compared to normal weight subjects including, but not limited to, drugs to treat atherosclerosis, osteoarthritis, disc herniation, degeneration of knees and hips, breast, endometrium, cervical, colon, leukemia and prostate cancers, hyperlipidemia, asthma/reactive airway disease, gallstones, GERD, obstructive sleep apnea, obesity hypoventilation syndrome, recurrent ventral hernias, menstrual irregularity and infertility.

Also provided herein are methods for treating obesity or obesity associated co-morbidities or other diseases or conditions involving K_(ATP) channels by co-administration to a subject in need thereof of an effective amount of any of the compounds according to Formulae I-VIII, or a salt of any of the compounds according to Formulae I-VIII or pharmaceutical formulation thereof, and a drug selected from the group consisting of an amphetamine or amphetamine mixture, Sibutramine, Orlistat, Rimonabant, a CB-1 antagonist, a 5HT_(2c) receptor agonist, a drug used to treat addiction, a beta adrenergic receptor agonist, an ACC inhibitor, leptin, a leptin analogue, a leptin agonist, a somatostatin agonist, an adiponectin agonist or secretagogue, Amylin, PYY or a PYY analogue, a ghrelin antagonist, a drug that inhibits gastrointestinal lipases or other digestive enzymes, a de-novo lipogenesis inhibitor, a drug that blocks absorption of dietary fat, growth hormone or a growth hormone analogue, a growth hormone secretagogue, a CCK agonist, an oleoylethanolamine receptor agonist, a fatty acid synthase inhibitor, a thyroid receptor agonist, a selective androgen receptor modulator, a PPAR agonist, a Beta hydroxysteroid Dehydrogenase-2 inhibitor, oxyntomodulin, oleoylestrone, a NPY2 receptor antagonist, a NPY5 receptor antagonist, a NPY agonist, a monoamine uptake inhibitor, a MTP inhibitor, a MC4 receptor agonist, a MCH1 receptor antagonist, a 5HT-6 antagonist, a histamine-3 antagonist, a glycine analog, a fgfl inhibitor, a DGAT-1 inhibitor, a carboxypeptidase inhibitor, an appetite suppressant, a non-thiazide diuretic, a drug that lowers cholesterol, a drug that raises HDL cholesterol, a drug that lowers LDL cholesterol, a drug that lowers blood pressure, a drug that is an anti-depressant, a drug that improves insulin sensitivity, a drug that improves glucose utilization or uptake, a drug that is an anti-epileptic, a drug that is an anti-inflammatory, a drug that is an appetite suppressant, a drug that lowers circulating triglycerides, a drug that is used to induce weight loss in an overweight or obese individual, and pharmaceutically acceptable salts thereof.

Provided herein are methods of (a) inhibiting or preventing the progression of type I diabetes, (b) reducing insulin dosing, (c) increasing glycemic control, or (d) delaying the loss of residual insulin secretion in a subject suffering from type I diabetes, comprising administering to said subject a therapeutically effective amount of a formulation of a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII. For example, the formulation may be selected from the group consisting of: i) a formulation comprising a salt, said salt comprising an anion of a K_(ATP) channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising a tertiary amine or ammonium group; ii) a formulation comprising a salt, said salt comprising an anion of a K_(ATP) channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII; and iii) a formulation comprising a salt, said salt comprising an anion of a K_(ATP) channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII, wherein at least one substituent comprises an amino group. In some embodiments of the methods, the formulation may be administered once, twice or three times per 24 hours. In certain embodiments, the formulation comprises diazoxide choline.

As noted above, the methods described herein can be used to inhibit or prevent the progression of Type I diabetes. In some such methods, the rate of beta cell loss is decreased by, e.g., about 5% to about 95% compared to the subject prior to administration of the formulations of the present invention. In other instances, the rate of beta cell loss is decreased by at least about 5%, 10%, 15%, 20%, 25%, 30% 40%, 50%, 60%, 70%, 80%, 90%, or 95% compared to the subject prior to administration of the formulations of the present invention, or prior to administration of a combination of the formulation as described herein and additional therapeutic agent(s).

The methods described herein can be used to reduce insulin dosing as described above. In some instance of the methods, the insulin dosing is decreased by about 5% to about 95% in a subject, compared to the subject's insulin dose prior to being administered the formulations of the present invention, or prior to administration of a combination of the formulation as described herein and additional therapeutic agent(s). In other instances of the methods, the insulin dosing is decreased by at least about 5%, 10%, 20%, 30%, or 50%.

As described above, the methods described herein can be used to increase glycemic control. “Glycemic control” refers to attempts to reproduce natural physiological glucose homeostasis. Accordingly, glycemic control is increased in a subject when the natural physiological glucose homeostasis is attained more often in a subject administered the formulations of the present invention compared to the subject prior to administration of the formulations.

The methods described herein can be used to delay the loss of residual insulin secretion. Residual insulin secretion refers to the partial preservation of islet beta-cell function, as demonstrated by preserved insulin production in subjects having Type I diabetes. Although not sufficient for the needs of the individual, residual insulin secretion is important for metabolic control, for avoidance of hypoglycemic episodes, and for protection against diabetic complications. To retain residual insulin secretion in type I diabetes is highly desirable. As used herein, the term “residual insulin secretion” refers to a level of insulin production by islet beta cells that is least about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or 60% of the level produced by a subject not diagnosed with Type I diabetes. In one instance, the methods provide that the loss of residual insulin secretion is delayed by at least about 3 months, 6 months, 12 months, 15 months, 18 months, 21 months, or up to 48 months compared to a subject not administered the formulations of the present invention.

Further provided herein are methods of increasing insulin sensitivity in a subject suffering from type II diabetes, comprising administering to said subject a therapeutically effective amount of a formulation of a KATP channel opener selected from the salts of the compounds of Formulae I-VIII. For example, the formulation may be selected from the group consisting of: i) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising a tertiary amine or ammonium group; ii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII; and iii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII, wherein at least one substituent comprises an amino group. In the methods, the therapeutically effective amount of the formulation may range from about 15 mg/day to about 500 mg/day. In some embodiments of the methods, the formulation may be administered once, twice or three times per 24 hours. In certain embodiments, the formulation comprises diazoxide choline.

The methods described herein can be used to increase insulin sensitivity in a subject by, e.g., about 10% to about 95% or more. In one instance, the insulin sensitivity is increased in a subject by at least about 30% compared to the subject prior to administration of the formulations of the present invention. In other instances, the insulin sensitivity is increased by at least about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80% or 90% compared to the subject prior to administration of the formulations of the present invention, or prior to administration of a combination of the formulation as described herein and additional therapeutic agent(s). Insulin sensitivity is indicated, for example, by reductions in the homeostasis model assessment-insulin resistance (HOMA-IR) index.

Still further provided are methods of treating a subject suffering from type II diabetes, comprising co-administering to said subject a therapeutically effective amount of a formulation and an anti-diabetic drug that either stimulates insulin secretion or is a short acting anti-diabetic drug, wherein the formulation is selected from the group consisting of: i) a formulation comprising a KATP channel opener selected from the group consisting of Formulae I-VIII; ii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising a tertiary amine or ammonium group; iii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII; and iv) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII, wherein at least one substituent comprises an amino group. In related embodiments of the above method, the anti-diabetic drug stimulates insulin secretion in a meal dependent fashion or is a short acting anti-diabetic drug with a circulating half life less than 5 hours. In the methods, the formulation and the anti-diabetic drug may be co-administered separately, sequentially or simultaneously. Suitable anti-diabetic drugs include but are not limited to exenatide, natiglinide, mitglinide, repaglinide, a DPP-IV inhibitor (for example vildagliptin, sitagliptin and saxagliptin), a PPAR agonist or partial agonist, a GLP-1 analog (e.g., liraglutide) or mimetic (e.g., modified exendin-4 analogue) or agonist, a thiazolidinedione, a disaccharidase inhibitor, a fructose-1,6-bisphosphatase inhibitor, a sulfonylurea receptor antagonist such as meglitinides and pharmaceutically acceptable salts thereof.

In some embodiments of the methods, the formulation may be administered once, twice or three times per 24 hours. In certain embodiments, the formulation comprises diazoxide choline.

In some embodiments of treating type II diabetes, the anti-diabetic drug may be one or more agents selected from the group consisting of a GLP-1 analog or mimetic, and a sulfonylurea receptor antagonist such as meglitinides.

Suitable sulfonylurea receptor antagonists for use in methods described herein include one or more agents selected from the group consisting of, but not limited to nateglinide, mitglinide, repaglinide and pharmaceutically acceptable salts thereof.

Suitable DPP-IV inhibitors for use in the present methods include one or more agents selected from the group consisting of, but limited to sitagliptin, saxagliptin, vildagliptin, and pharmaceutically acceptable salts thereof.

GLP-1 analogs or mimetics that may be used in the present methods include one or more agents selected from the group consisting of exenatide, liraglutide, albugon, exendin-4 analogs and conjugates, and pharmaceutically acceptable salts and conjugates thereof.

Suitable meglitinides for use in the present methods include one or more agents selected from the group consisting of, but not limited to repaglinide, nateglinide, mitiglinide and pharmaceutically acceptable salts thereof.

Likewise suitable thiazolidinediones, PPAR agonists or partial agonists that may be used in the methods described herein include one or more agents selected from the group consisting of, but not limited to pioglitazone, rosiglitazone, troglizazone, tesaglitazar, muraglitazar, netoglitazar, LY518674, ragaglitazar, sipoglitazar, and pharmaceutically acceptable salts thereof.

Exemplary disaccharidase inhibitor for use in the present methods include one or more agents selected from the group consisting of, but not limited to acarbose, AO-128, and pharmaceutically acceptable salts thereof.

Fructose-1,6-bisphosphatase inhibitors suitable for use in the present methods include one or more agents selected from the group consisting of, but not limited to phenyl phosphonates, benzimidazoles, CS-917, anilinoqiunazolones, and benzoxaozole benzenesulfonamides, and pharmaceutically acceptable salts thereof.

The formulation and anti-diabetic may be combined in a single pharmaceutical composition. For example, the formulation and anti-diabetic may be combined in a controlled release pharmaceutical composition comprising 50 to 500 mg of diazoxide choline and 20 to 100 mg of sitagliptin. While typically such controlled release pharmaceutical compositions are administered once per 24 hours, other dosing regimens are contemplated including two or three times a day or every other day.

Exemplary anti-diabetic drug for use in the present methods include one or more agents selected from the group consisting of, exenatide, nateglinide, mitglinide, repaglinide, and pharmaceutically acceptable salts or conjugates thereof.

Alternatively, the formulation may comprise 50 to 450 mg of diazoxide choline and is administered once per 24 hours as a controlled release oral formulation, and the anti-diabetic is a pharmaceutical composition comprising 5 to 10 μg exenatide and is administered twice per 24 hours, before meals. In such methods, the controlled release oral formulation may be a tablet and the exenatide may be administered by subcutaneous injection.

In some embodiments of methods of treating type II diabetes, the formulation comprises 50 to 500 mg of diazoxide choline and is administered once per 24 hours as a controlled release oral formulation, and the anti-diabetic is a pharmaceutical composition comprising the meglitinide and is administered three times per 24 hours, before meals.

Provided herein are methods of treating a dyslipidemia by (a) reducing an abnormally high total cholesterol level in a subject's blood, (b) reducing an abnormally high LDL cholesterol level in a subject's blood, (c) reducing an abnormally high VLDL cholesterol level in a subject's blood (d) reducing an abnormally high non-HDL cholesterol level in a subject's blood (e) reducing an abnormally high triglyceride level in a subject's blood and/or (b) raising an abnormally low HDL cholesterol level in a subject's blood level, comprising administering to said subject a therapeutically effective amount of a formulation selected from the group consisting of:

i) a formulation comprising a K_(ATP) channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, Formula V, Formula VI, Formula VII, and Formula VIII;

ii) a formulation comprising a salt, said salt comprising an anion of a K_(ATP) channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising a tertiary amine or ammonium group;

iii) a formulation comprising a salt, said salt comprising an anion of a K_(ATP) channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII; and

iv) a formulation comprising a salt, said salt comprising an anion of a K_(ATP) channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII, wherein at least one substituent comprises an amino group. As used herein, a reduction in the various specified cholesterol containing lipoproteins in the circulation amounts to a reduction, e.g., by about 1% to about 60%, by about 1% to about 30%, by about 2% to about 20%. In other embodiments the subject's HDL cholesterol level may be raised, e.g., by about 1% to about 60%, by about 1% to about 50%, by about 2% to about 40%. The subject's triglycerides level may also be reduced by the present methods by about 5% to about 95%, by about 5% to about 90%, by about 5% to about 85%, by about 10% to about 95%, by about 10% to about 90%, or by about 10% to about 85%. The subject may suffer from or may be at risk for pancreatitis and or may be obese. In some embodiments the formulation is administered once, twice or three times per 24 hours. In other embodiments, the formulation comprises diazoxide choline.

In some embodiments, multiple administrations of the formulation are given over a period of days. In such cases, the caloric intake of the subject during the period of days is substantially the same as before the formulation was administered. As used in this context, “substantially” means less than 15% change, more preferably less than 10% change, or even more preferably at least less than 5% change. In a most preferred embodiment, the caloric intake of the subject is the same before administration and during the period of days during administration.

Provided herein are methods of (a) reducing circulating androgen levels, or (b) reestablishing normal ovulation cycles, in a subject suffering from poly-cystic ovarian syndrome, comprising administering to said subject a therapeutically effective amount of a formulation of a KATP channel opener selected from the salts of the compounds of Formulae I-VIII. For example, the formulation may be selected from the group consisting of: i) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising a tertiary amine or ammonium group; ii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII; and iii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII, wherein at least one substituent comprises an amino group. In some embodiments of the methods, the formulation may be administered once, twice or three times per 24 hours. In certain embodiments, the formulation comprises diazoxide choline.

As noted above, methods described herein can be used to reduce the circulating androgen levels in a subject by, e.g., about 5% to about 95% of the level of circulating androgen compared to the subject prior to administration of the formulations of the present invention. In other instances, the level of circulating androgen is reduced by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70% or 80% compared to the subject prior to administration of the formulations of the present invention, or prior to administration of a combination of the formulation as described herein and additional therapeutic agent(s). Circulating androgen can be measured in a subject using any suitable method known in the art.

The methods described herein can be used to reestablish normal ovulation cycles in a subject. Ovulation is the time in a female subject's menstrual cycle when the ovum, or egg, is released. It is when a woman is most fertile and likely to conceive. Ovulation occurs 12 or 14 days before menstruation. A menstruation period begins about every 28 days if the woman does not become pregnant in a given cycle. In one instance, a normal ovulation cycle of approximately 28 days is reestablished in a subject, where the subject experienced a cycle of greater than or fewer than 28 days prior to administration of the formulations of the present invention. Ovulation can be measured using any means available, including urinary kits, basal body temperature tests, a cervical mucus test and a blood test.

Still further provided are methods of treating a subject suffering from poly-cystic ovarian syndrome comprising co-administering to said subject a therapeutically effective amount of a formulation and an anti-androgen therapy, wherein the formulation is selected from the group consisting of: i) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising a tertiary amine or ammonium group; ii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII; and iii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII, wherein at least one substituent comprises an amino group. In the methods, the formulation and the anti-androgen therapy may be co-administered separately, sequentially or simultaneously. In some embodiments of the methods, the formulation may be administered once, twice or three times per 24 hours. In certain embodiments, the formulation comprises diazoxide choline.

Also provided are methods of treating a subject suffering from poly-cystic ovarian syndrome comprising co-administering to said subject a therapeutically effective amount of a formulation and a selective estrogen receptor modulator (SERM) therapy, wherein the formulation is selected from the group consisting of: i) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising a tertiary amine or ammonium group; ii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII; and iii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII, wherein at least one substituent comprises an amino group. In the methods, the formulation and the selective estrogen receptor modulator (SERM) therapy may be co-administered separately, sequentially or simultaneously. In some embodiments of the methods, the formulation may be administered once, twice or three times per 24 hours. In certain embodiments, the formulation comprises diazoxide choline. In some embodiments of the methods the SERM includes clomiphene (or a salt thereof) or ormeloxifene.

Further provided are methods of treating a subject suffering from poly-cystic ovarian syndrome comprising co-administering to said subject a therapeutically effective amount of a formulation and an ovulation inducing therapy, wherein the formulation is selected from the group consisting of: i) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising a tertiary amine or ammonium group; ii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII; and iii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII, wherein at least one substituent comprises an amino group. In the methods, the formulation and the ovulation inducing therapy may be co-administered separately, sequentially or simultaneously. In some embodiments of the methods, the formulation may be administered once, twice or three times per 24 hours. In certain embodiments, the formulation comprises diazoxide choline. In some embodiments of the methods the ovulation inducing therapy includes follicle stimulating hormone.

Still further provided are methods of treating a subject suffering from poly-cystic ovarian syndrome comprising co-administering to said subject a therapeutically effective amount of a formulation and an aromatase inhibitor therapy, wherein the formulation is selected from the group consisting of: i) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising a tertiary amine or ammonium group; ii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII; and iii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII, wherein at least one substituent comprises an amino group. In the methods, the formulation and the aromatase inhibitor therapy may be co-administered separately, sequentially or simultaneously. In some embodiments of the methods, the formulation may be administered once, twice or three times per 24 hours. In certain embodiments, the formulation comprises diazoxide choline. In some embodiments of the methods the aromatase inhibitor therapy includes letrozole or anastrozole.

Provided herein are methods of treating a subject suffering from hypertension comprising administering to said subject a therapeutically effective amount of a single oral agent wherein the oral agent is a formulation of a KATP channel opener selected from the salts of the compounds of Formulae I-VIII. For example, the formulation may be selected from the group consisting of: i) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising a tertiary amine or ammonium group; ii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII; and iii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII, wherein at least one substituent comprises an amino group. In some embodiments of the methods, the formulation may be administered once, twice or three times per 24 hours. In certain embodiments, the formulation comprises diazoxide choline.

Also provided herein are methods of treating a subject suffering from hypertension comprising co-administering to said subject a therapeutically effective amount of a formulation and an oral anti-hypertensive agent, wherein the formulation of a KATP channel opener is selected from the salts of the compounds of Formulae I-VIII. For example, the formulation may be selected from the group consisting of: i) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising a tertiary amine or ammonium group; ii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII; and iii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII, wherein at least one substituent comprises an amino group. In some embodiments of the methods, the formulation may be administered once, twice or three times per 24 hours. In certain embodiments, the formulation comprises diazoxide choline.

Further provided are methods of treating a subject suffering from pulmonary hypertension comprising administering to said subject a therapeutically effective amount of a formulation selected from the group consisting of:

i) a formulation comprising a salt, said salt comprising an anion of a K_(ATP) channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising a tertiary amine or ammonium group;

ii) a formulation comprising a salt, said salt comprising an anion of a K_(ATP) channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII; and

iii) a formulation comprising a salt, said salt comprising an anion of a K_(ATP) channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII, wherein at least one substituent comprises an amino group. The amount of the formulation is effective to lower pulmonary arteriolar resistance and pulmonary artery pressure and, in some embodiments, to bring pulmonary arteriolar resistance and pulmonary artery pressure within normal limits (systolic 15-30 mm Hg, diastolic 6-12 mm Hg). Treatment may be continued as long as the subject continues to suffer from pulmonary hypertension. In some embodiments of the methods, the formulation may be administered once, twice or three times per 24 hours. In certain embodiments, the formulation comprises diazoxide choline, in the form of, e.g., controlled release tablets.

Also provided herein are methods of treatment to prevent late post-myocardial infarction arrhythmias comprising administering to a subject that has recently suffered a myocardial infarction a therapeutically effective amount of a pharmaceutical formulation wherein the formulation includes a K_(ATP) channel opener selected from the salts of the compounds of Formulae I-VIII. For example, the formulation may be selected from the group consisting of:

i) a formulation comprising a salt, said salt comprising an anion of a K_(ATP) channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising a tertiary amine or ammonium group;

ii) a formulation comprising a salt, said salt comprising an anion of a K_(ATP) channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII; and

iii) a formulation comprising a salt, said salt comprising an anion of a K_(ATP) channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII, wherein at least one substituent comprises an amino group. By recently suffered a myocardial infarction is meant that the myocardial infarction has occurred within the last 48 hours. The subject is typically treated for a minimum of one year or until the period of late remodeling is complete. In some embodiments of the methods, the formulation may be administered once, twice or three times per 24 hours. In certain embodiments, the formulation comprises diazoxide choline, in the form of, e.g., controlled release tablets.

Also provided herein are methods of treating a subject immediately following the detection of myocardial infarction to limit infarct size and prevent arrhythmias. The methods include administering to the subject a therapeutically effective amount of a pharmaceutical formulation selected from the group consisting of:

i) a formulation comprising a salt, said salt comprising an anion of a K_(ATP) channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising a tertiary amine or ammonium group;

ii) a formulation comprising a salt, said salt comprising an anion of a K_(ATP) channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII; and

iii) a formulation comprising a salt, said salt comprising an anion of a K_(ATP) channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII, wherein at least one substituent comprises an amino group. Treatment may be continued after the infarction for a minimum of one year or until the period of late remodeling is complete. In some embodiments of the methods, the formulation may be administered once, twice or three times per 24 hours. In certain embodiments, the formulation comprises diazoxide choline, in the form of, e.g., controlled release tablets. As used herein, reference to beginning treatment “immediately” following the detection of myocardial infarction means that the treatment is begun within one hour after diagnosis of infarction, and less preferably between 1-2 hours, 2-3, 3-4 hours, 4-8 hours and 8-24 hours post diagnosis of infarction.

Provided herein are methods of treating a subject at risk for myocardial infarction to prevent infarction, limit infarct size, and/or to prevent arrhythmias. The methods include administering to the subject a therapeutically effective amount of a pharmaceutical formulation selected from the group consisting of:

i) a formulation comprising a salt, said salt comprising an anion of a K_(ATP) channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising a tertiary amine or ammonium group;

ii) a formulation comprising a salt, said salt comprising an anion of a K_(ATP) channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII; and

iii) a formulation comprising a salt, said salt comprising an anion of a K_(ATP) channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII, wherein at least one substituent comprises an amino group. Treatment may be continued for as long as the subject is at risk for myocardial infarction. In some embodiments of the methods, the formulation may be administered once, twice or three times per 24 hours. In certain embodiments, the formulation comprises diazoxide choline, in the form of, e.g., controlled release tablets.

In the above methods relating to treating heart disease such as myocardial infarction (or its prevention) or arrhythmias, the administration of the KATP channel opener (such as diazoxide) may be beneficial by acting as an anti-inflammatory agent. Diazoxide is known to have anti-inflammatory effects (see Xu et al., J. Hypertension, 2006 24: 915-922; Wang et al., Shock 2004, 22:23-28).

Provided herein are methods of treating a subject suffering from hypoglycemia-associated autonomic failure comprising administering to said subject a therapeutically effective amount of a formulation of a KATP channel opener selected from the salts of the compounds of Formulae I-VIII. For example, the formulation may be selected from the group consisting of: i) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising a tertiary amine or ammonium group; ii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII; iii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII, wherein at least one substituent comprises an amino group; and iv) a formulation comprising 7-chloro-3-methyl-2H-1,2,4-benzothiadiazine. In the methods the subject may also be suffering from type I diabetes or type II diabetes. In some embodiments of the methods, the formulation may be administered once, twice or three times per 24 hours. The formulation may also be an oral or an intranasal formulation. In certain embodiments, the formulation comprises diazoxide choline.

A method of inducing or increasing beta-cell rest in a subject in need thereof, comprising administering to said subject a therapeutically effective amount of a formulation of a KATP channel opener selected from the salts of the compounds of Formulae I-VIII. For example, the formulation may be selected from the group consisting of: i) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising a tertiary amine or ammonium group; ii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII; and iii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII, wherein at least one substituent comprises an amino group. In some embodiments of the methods, the formulation may be administered once, twice or three times per 24 hours. In certain embodiments, the formulation comprises diazoxide choline. In the methods, the subject may also be obese or overweight or suffer from type II diabetes.

Also provided herein are polymorphic forms (i.e., “polymorphs”) of the compounds of Formulae I-VIII, as exemplified by the X-ray Power Diffraction (XRPD) patterns shown in any of the figures.

Also provided are polymorphs of salts of diazoxide which include diazoxide and a cation selected from the group consisting of an alkali metal and a compound comprising a tertiary amine or quaternary ammonium group.

Also provided herein are methods for producing a diazoxide choline salt, which includes suspending diazoxide in a solvent and mixing with a choline salt, adding a co-solvent to the suspension under conditions sufficient to cause formation and precipitation of the diazoxide choline salt, and harvesting the precipitate to provide the diazoxide choline salt.

Also provided herein are methods of treating obesity or obesity-related co-morbidity in an obese subject, wherein the method comprising administering to a subject in need thereof an effective amount of a compound of Formula V-VIII.

Also provided herein are methods for treatment of a subject suffering from or at risk for Alzheimer's disease (AD), which methods include administration to a subject a therapeutically effective amount of a pharmaceutical formulation comprising a compound of any of Formulae I-VIII as provided herein. In some embodiments, the compound is diazoxide. Also provided herein are methods for treatment of a subject suffering from or at risk for AD, which methods include administration to a subject a therapeutically effective amount of a salt of a compound according to any of Formulae I-VIII. In some embodiments, the compound is a salt of diazoxide.

In another embodiment, the invention provides a method for treating hypoglycemia by administration of an effective amount of a pharmaceutical formulation comprising a salt selected from the group consisting of a) a salt comprising an anion of a KATP channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising a tertiary amine or ammonium group; b) a salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII; and c) a salt comprising a cation of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII, wherein at least one substituent comprises an amino group.

In further embodiments, the hypoglycemia is selected from the group consisting of a) nighttime hypoglycemia, b) hypoglycemia attributable to a defect in insulin secretion, c) attributable to an insulin secreting tumor, and d) drug-induced hypoglycemia.

Provided herein are methods of treating obesity or type 1 or type 2 diabetes in a companion animal. The methods include administering to said animal a therapeutically effective amount of a formulation selected from the group consisting of: i) a formulation comprising a KATP channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, Formula V, Formula VI, Formula VII, and Formula VIII; ii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising a tertiary amine or ammonium group; iii) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII; and iv) a formulation comprising a salt, said salt comprising an anion of a KATP channel opener selected from the group consisting of Formula V, Formula VI, Formula VII and Formula VIII, wherein at least one substituent comprises an amino group. Typically, the animal is a cat, a dog or a horse. The formulation can be suitable for oral administration, e.g., a controlled release tablet or capsule, or a chewable formulation. Alternatively, the formulation may be a granulated controlled release formulation which can be applied to food for the animal, e.g., a powder. In some embodiments, the formulation is administered once or twice per 24 hours. In certain embodiments, the formulation comprises diazoxide choline.

Provided herein are methods of treating a dyslipidemia in a subject having a triglyceride level of at least about 500 mg/dL and an HDL-C level of about 40 mg/dL or less, the method comprising administering to the subject a therapeutically effective amount of a K_(ATP) channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and salts thereof. In some embodiments, the K_(ATP) channel opener comprises an anion Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising an ammonium group comprising at least one tertiary amine group. In some embodiments, the said K_(ATP) channel opener comprises a salt of diazoxide; such as diazoxide choline. In some embodiments, the K_(ATP) channel opener is formulated as a controlled release formulation for oral administration; such as a formulation formulated for once or twice a day administration. In some embodiments, the subject is administered about 145 to 435 mg per day of said K_(ATP) channel opener. In some embodiments, the K_(ATP) channel opener is co-administered or co-formulated with a therapeutically effective amount of a second active compound for lowering triglycerides, raising HDL-C, lowering LDL-C, or any combination thereof. In some related embodiments, the second active compound comprises a statin or a fibrate. In some related embodiments, the second active compound comprises a fibrate or a salt thereof; such as fenofibrate or a salt thereof; such as fenofibrate choline. In some further related embodiments, the subject is administered about 45 to 200 mg per day of fenofibrate choline. In some embodiments, the subject is a human.

Provided herein are methods for treating dyslipidemia in a subject having a triglyceride level of at least about 1000 mg/dL, the methods comprising administering to the subject a therapeutically effective amount of a fibrate or a salt thereof and a therapeutically effective amount of a K_(ATP) channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and salts thereof. In some embodiments, the K_(ATP) channel opener comprises an anion Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising an ammonium group comprising at least one tertiary amine group. In some embodiments, the said K_(ATP) channel opener comprises a salt of diazoxide; such as diazoxide choline. In some embodiments, the K_(ATP) channel opener is formulated as a controlled release formulation for oral administration; such as a formulation formulated for once or twice a day administration. In some embodiments, the subject is administered about 145 to 435 mg per day of said K_(ATP) channel opener. In some embodiments, the fibrate or a salt thereof; such as fenofibrate or a salt thereof; such as fenofibrate choline. In some related embodiments, the subject is administered about 45 to 200 mg per day of fenofibrate choline. In some embodiments, the fibrate or a salt thereof and the K_(ATP) channel opener are co-formulated, such as a controlled release formulation for oral administration, such as formulated for once or twice a day administration. In some embodiments, the subject is human.

Provided herein are methods for reducing elevated triglycerides in a subject undergoing statin therapy, the methods comprising administering to the subject a therapeutically effective amount of a K_(ATP) channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and salts thereof. In some embodiments, the K_(ATP) channel opener comprises an anion Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising an ammonium group comprising at least one tertiary amine group. In some embodiments, the said K_(ATP) channel opener comprises a salt of diazoxide; such as diazoxide choline. In some embodiments, the K_(ATP) channel opener is formulated as a controlled release formulation for oral administration; such as a formulation formulated for once or twice a day administration. In some embodiments, the subject is administered about 145 to 435 mg per day of said K_(ATP) channel opener. In some embodiments, the K_(ATP) channel opener is co-administered or co-formulated with a therapeutically effective amount of a third active compound for lowering triglycerides. In some related embodiments, the third active compound comprises a fibrate or a salt thereof; such as fenofibrate or a salt thereof; such as fenofibrate choline. In some further related embodiments, the subject is administered about 45 to 200 mg per day of fenofibrate choline. In some related embodiments, the fibrate or a salt thereof and the K_(ATP) channel opener are co-formulated, such as a controlled release formulation for oral administration, such as for once or twice a day administration. In some related embodiments, the subject is human.

Provided herein are methods for treating a subject with nonalcoholic steatohepatitis (NASH), the methods comprising administering to the subject a therapeutically effective amount of a K_(ATP) channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and salts thereof. In some embodiments, the K_(ATP) channel opener comprises an anion Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising an ammonium group comprising at least one tertiary amine group. In some embodiments, the said K_(ATP) channel opener comprises a salt of diazoxide; such as diazoxide choline. In some embodiments, the K_(ATP) channel opener is formulated as a controlled release formulation for oral administration; such as a formulation formulated for once or twice a day administration. In some embodiments, the subject is administered about 145 to 435 mg per day of said K_(ATP) channel opener. In some embodiments, the K_(ATP) channel opener is co-administered or co-formulated with a therapeutically effective amount of a second active compound. In some related embodiments, the second active compound comprises a fibrate or a salt thereof; such as fenofibrate or a salt thereof; such as fenofibrate choline. In some further related embodiments, the subject is administered about 45 to 200 mg per day of fenofibrate choline. In some related embodiments, the second active compound comprises a statin. In some related embodiments, the second active compound and the K_(ATP) channel opener are co-formulated, such as a controlled release formulation for oral administration, such as for once or twice a day administration. In some related embodiments, the subject is human.

Also provided herein are pharmaceutical formulations comprising diazoxide choline, wherein oral administration of a 290 mg dose of diazoxide choline in the formulations once daily to a subject results in steady state AUC₀₋₂₄>500 μg*hr/mL. In some embodiments, oral administration of a 290 mg dose of diazoxide choline in the formulations once daily to a subject further results in steady state % Peak-to-Trough Fluctuation of less than 30%. In some embodiments, oral administration of a 290 mg dose of diazoxide choline in the formulations once daily to a subject results in steady state average circulating drug level (C_(av(ss))) between 14 and 31 μg/mL. In some embodiments, the pharmaceutical formulations are compressed tablet formulations.

Also provided herein are pharmaceutical formulations comprising a salt of diazoxide formulated for oral administration, wherein bioavailability of diazoxide is at least about 50%; such as at least about 75%; such as at least about 90%. In some related embodiments, the salt of diazoxide is diazoxide choline. In some further related embodiments, diazoxide choline is of polymorph form B. In some embodiments, the pharmaceutical formulations are compressed tablet formulations.

Also provided herein are methods for treating a dyslipidemia in a subject, the method comprising administering to the subject: a) a formulation comprising one or more omega-3 fatty acids, and b) a formulation comprising a therapeutically effective amount of a K_(ATP) channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and salts thereof. In some embodiments, the K_(ATP) channel opener comprises an anion Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising an ammonium group comprising at least one tertiary amine group. In some embodiments, the dyslipidemia is selected from the group consisting of abnormally high total cholesterol level in a subject's blood, abnormally high LDL cholesterol, abnormally high VLDL cholesterol, abnormally high non-HDL cholesterol, abnormally high triglycerides, abnormally low HDL cholesterol, or any combination thereof. In some embodiments, the dyslipidemia comprises abnormally high triglycerides; such as a triglyceride level of at least about 500 mg/dL; such as a triglyceride level of at least about 1000 mg/dL. In some embodiments, the said K_(ATP) channel opener comprises a salt of diazoxide; such as diazoxide choline. In some embodiments, the K_(ATP) channel opener is formulated as a controlled release formulation for oral administration; such as a formulation formulated for once or twice a day administration. In some embodiments, the subject is administered about 145 to 435 mg per day of said K_(ATP) channel opener. In some embodiments, the omega-3 fatty acid formulation is substantially free of DHA.

Also provided herein are methods for initiating treatment for a dyslipidemia in a subject, the methods comprising administering a fibrate or a salt thereof and a K_(ATP) channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and salts thereof; wherein the fibrate or a salt thereof and the K_(ATP) channel opener are both initially administered at sub-optimal titration doses, and the doses of both are increased to maintenance doses over 7 to 28 days. In some embodiments, the K_(ATP) channel opener comprises an anion Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising an ammonium group comprising at least one tertiary amine group. In some embodiments, the dyslipidemia comprises a triglyceride level of at least about 500 mg/dL. In some related embodiments, the dyslipidemia further comprises a HDL-C level of less than about 40 mg/dL. In some embodiments, the dyslipidemia comprises a triglyceride level of at least about 1000 mg/dL. In some embodiments, the titration doses of the fibrate or a salt thereof and the K_(ATP) channel opener are about between about one third to about one half of the corresponding maintenance doses. In some embodiments, the K_(ATP) channel opener is formulated as a controlled release formulation for oral administration for oral administration; such as for once or twice daily administration. In some embodiments, the K_(ATP) channel opener comprises a salt of diazoxide; such as diazoxide choline. In some embodiments, the K_(ATP) channel opener titration dose is about 70 mg to 220 mg. In some embodiments, the K_(ATP) channel opener maintenance dose is about 145 mg to 435 mg. In some embodiments, the fibrate or a salt thereof is formulated as a controlled release formulation for oral administration; such as for once or twice daily administration. In some embodiments, the fibrate fibrate or a salt thereof comprises fenofibrate or a salt thereof; such as fenofibrate choline. In some embodiments, the titration dose of the fibrate or a salt thereof is about 35 mg to 100 mg. In some embodiments, the maintenance dose of the fibrate or a salt thereof is about 100 mg to about 200 mg. In some embodiments, the titration doses of the fibrate or a salt thereof and the K_(ATP) channel opener are co-formulated; such as co-formulated as a controlled release formulation for oral administration; such as for once or twice daily administration. In some embodiments, the maintenance doses of the fibrate or a salt thereof and the K_(ATP) channel opener are co-formulated; such as co-formulated as a controlled release formulation for oral administration; such as for once or twice daily administration.

Also provided herein are kits comprising: a pharmaceutical treatment blister card with a plurality of blister cavities; seven to twenty eight daily dosages of a K_(ATP) channel opener; and seven to twenty eight daily dosages of a fibrate or a salt thereof. In these kits, the daily dosages of the K_(ATP) channel opener and the fibrate or a salt thereof are sub-optimal titration dosages. In some embodiments, each of the plurality of blister cavities contain a pharmaceutical formulation comprising either a K_(ATP) channel opener or a fibrate or a salt thereof. In some embodiments, each of the plurality of blister cavities contains two or more pharmaceutical formulations, wherein at least one of said pharmaceutical formulations comprises a K_(ATP) channel opener, and at least one other of said pharmaceutical formulations comprises a fibrate or a salt thereof. In some embodiments, the K_(ATP) channel opener and said fibrate are co-formulated in a single pharmaceutical formulation. In some related embodiments, each of the plurality of blister cavities contains the co-formulated pharmaceutical formulation. In some embodiments, the K_(ATP) channel opener is selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and salts thereof. In some embodiments, the K_(ATP) channel opener comprises an anion Formula I, Formula II, Formula III and Formula IV, and a cation selected from the group consisting of an alkali metal and a compound comprising an ammonium group comprising at least one tertiary amine group. In some embodiments, the K_(ATP) channel opener comprises a salt of diazoxide; such as diazoxide choline. In some embodiments, the fibrate or a salt thereof comprises fenofibrate or a salt thereof; such as fenofibrate choline.

Also provided herein are methods for treating a subject with acute pancreatitis, the method comprising intravenously administering a bolus dose of a K_(ATP) channel opener of between about 0.5 to 5 mg/kg over 5 to 10 minutes; wherein the K_(ATP) channel opener is selected from the group consisting of Formula I, Formula II, Formula III and Formula IV, and wherein the K_(ATP) channel opener is formulated for intravenous administration. In some embodiments, the dose of K_(ATP) channel opener administered as a bolus is between about 50 to 200 mg. In some embodiments, the methods further comprise additional administration of the K_(ATP) channel opener after completion of the bolus at between about 0.02 to 0.2 mg/kg/hr, along with administration of insulin on a recurring basis at a level and rate sufficient to maintain fasting glucose between about 70 mg/dL and 125 mg/dL and peak post prandial glucose less than 200 mg/dL. In some related embodiments, the insulin comprises a basal insulin, a fast-acting prandial insulin, or a mixture thereof. In some embodiments, the K_(ATP) channel opener comprises diazoxide or an ion thereof. In some embodiments, the methods further comprise co-administration of nicorandil. In some embodiments, the methods further comprise treating the subject using plasmapheresis. In some embodiments, the subject is a human.

In the present context, the term “therapeutically effective” or “effective amount” indicates that the materials or amount of material is effective to prevent, alleviate, or ameliorate one or more symptoms of a disease or medical condition, and/or to prolong the survival of the subject being treated.

The term “pharmaceutically acceptable” indicates that the identified material does not have properties that would cause a reasonably prudent medical practitioner to avoid administration of the material to a patient, taking into consideration the disease or conditions to be treated and the respective route of administration. For example, it is commonly required that such a material be essentially sterile, e.g., for injectibles.

As used herein, the term “composition” refers to a formulation suitable for administration to an intended animal subject for therapeutic purposes that contains at least one pharmaceutically active compound and at least one pharmaceutically acceptable carrier or excipient. Other terms as used herein are defined below.

As used herein, the term “co-administer” or “co-administration” refers to administration of two or more pharmaceutically active materials to a subject at the same time. For example, two compositions, each in tablet form, can be co-administered by oral ingestion at the same time or within minutes of each other. Co-administration also encompasses administration of two active materials co-formulated in a single delivery form. That is, two compositions can be combined in a single formulation (co-formulated) for administration to a subject so that the compositions are co-administered, e.g. in a single tablet, capsule, oral suspension, injectible liquid, etc.

Adipocyte: An animal connective tissue cell specialized for the synthesis and storage of fat.

Agonist: A chemical compound that has affinity for and stimulates physiological activity at cell receptors normally stimulated by naturally occurring substances, triggering a biochemical response. An agonist of a receptor can also be considered an activator of the receptor.

About: is used herein to mean in quantitative terms plus or minus 10%.

Adipose tissue: Tissue comprised principally of adipocytes.

Adolescent: A person between 10 and 19 years of age.

Adiponectin: A protein hormone produced and secreted exclusively by adipocytes that regulates the metabolism of lipids and glucose. Adiponectin influences the body's response to insulin. Adiponectin also has anti-inflammatory effects on the cells lining the walls of blood vessels.

Alkali metal: refers to elements included in Group I of the periodic table, such as, lithium, sodium, potassium, rubidium, cesium and francium.

Amelioration of the symptoms of a particular disorder by administration of a particular pharmaceutical composition: refers to any lessening, whether permanent or temporary, lasting or transient that can be attributed to or associated with administration of the composition.

Analog: a compound that resembles another in structure but differs by at least one atom.

Antagonist: A substance that tends to nullify the action of another, as a drug that binds to a cell receptor without eliciting a biological response when confronted with an agonist for the receptor.

Anti-androgen therapy: a therapy using medications to block production of or interfere with the action of male sex hormones, e.g. testosterone. Anti-androgen therapies suitable for use in the methods described herein include but are not limited to Progestin, cyproterone, ethinylestradiol, cyproterone, and androgen receptor protein inhibitors.

Atherosclerotic Plaque: A buildup of cholesterol and fatty material within a blood vessel due to the effects of atherosclerosis

Bariatric Surgery: A range of surgical procedures which are designed to aid in the management or treatment of obesity and allied diseases.

Beta cell rest: Temporarily placing beta cells in a condition in which there is reduced metabolic stress due to suppressed secretion of insulin.

Bilaminate: A component of a pharmaceutical dosage form that consists of the lamination of two distinct materials.

Bioavailability: Refers to the amount or extent of therapeutically active substance that is released from the drug product and becomes available in the body at the intended site of drug action. The amount or extent of drug released can be established by the pharmacokinetic-parameters, such as the area under the blood or plasma drug concentration-time curve (AUC) and the peak blood or plasma concentration (C max) of the drug.

Bioequivalent: Two formulations of the same active substance are bioequivalent when there is no significant difference in the rate and extent to which the active substance becomes available at the site of drug action when administered at the same molar dose under similar conditions. “Formulation” in this definition may include the free base of the active substance or different salts of the active substance. Bioequivalence may be demonstrated through several in vivo and in vitro methods. These methods, in descending order of preference, include pharmacokinetic, pharmacodynamic, clinical and in vitro studies. In particular, bioequivalence is demonstrated using pharmacokinetic measures such as the area under the blood or plasma drug concentration-time curve (AUC) and the peak blood or plasma concentration (Cmax) of the drug, using statistical criteria.

Cannabinoid Receptor: Receptors in the endocannabinoid (EC) system associated with the intake of food and tobacco dependency. Blocking the cannabinoid receptor may reduce dependence on tobacco and the craving for food.

Capsule: refers to a softgel, caplet, or any other encapsulated dosage form known to practitioners in the art, or a portion thereof. Softgel refers a soft gelatin capsule, in agreement with the accepted nomenclature adopted by the SoftGel Association. A softgel is a one-piece, sealed, soft gelatin (or other film-forming material) shell that contains a solution, a suspension, or a semi-solid paste.

Combination: Refers to any association between or among two or more items. The combination can be two or more separate items, such as two compositions or two collections. It can be a mixture thereof, such as a single mixture of the two or more items, or any variation thereof.

Combined hyperlipidemia: Refers to a commonly occurring form of hypercholesterolemia (elevated cholesterol level) characterized by increased LDL cholesterol and triglyceride concentrations, often accompanied by decreased HDL cholesterol.

Companion animal. Animals kept for the purpose of providing companionship to humans, e.g., house pets, including cats and dogs and horses.

Compression tablet: Tablet formed by the exertion of pressure to a volume of tablet matrix in a die.

Compression coated tablet: A tablet formed by the addition of a coating by compression to a compressed core containing the pharmaceutical active. As used herein the term “tablet” is intended to mean the same as a compression tablet unless indicated otherwise.

Derivative: A chemical substance derived from another substance by modification or substitution.

Daily dosage: The total amount of a drug taken in a 24 hour period whether taken as a single dose or taken in multiple doses.

DPP-IV inhibitor: A class of anti-diabetic drugs which inhibit the enzyme dipeptidylpeptidase-IV, also known as DPP-IV. The DPP-IV enzyme cleaves the N-terminal dipeptide from GLP-1, inactivating it. DPP-IV inhibitors thus increase the potency of GLP-1 biological effects.

Disaccharidase inhibitor: A class of anti-diabetic drugs which act by inhibiting disaccharidase enzymes. Disaccharidases break down disaccharides into monosaccharides and include, e.g., lactase, sucrase, trehalase, and maltase. Inhibition of disaccharidases delays and decreases absorption of monosaccharides by the body, decreasing post-meal glucose peaks.

Diazoxide: 7-chloro-3-methyl-2H-1,2,4-benzothiadiazine 1,1 dioxide (shown below with its tautomer) with the empirical formula C8H7ClN2O2S and a molecular weight of 230.7.

Dyslipidemia: A disorder of lipoprotein metabolism, including lipoprotein overproduction or deficiency. Dyslipidemias may be manifested by elevation of the total cholesterol, low-density lipoprotein (LDL) cholesterol and triglyceride, and a decrease in high-density lipoprotein (HDL) cholesterol concentration in the blood. Optimal LDL cholesterol level for adults is less than 100 mg/dL (2.60 mmol/L), optimal HDL cholesterol level is equal to or greater than 40 mg/dL (1.02 mmol/L), and optimal triglyceride level is less than 150 mg/dL (1.7 mmol/L). As used herein, a dyslipidemia is defined as an abnormal level which is at least 5% greater (or lower) than the normal level, more preferably at least 10% greater (or lower) than the normal level, more preferably at least 20% greater or lower than the normal level. Dyslipidemia is a risk factor for cardiovascular disease, predisposing affected individuals to atherosclerosis. Obese and obese diabetic patients experience higher rates of dyslipidemia. Although diazoxide administration has been reported to reduce circulating triglyceride levels in treated obese patients (Alemzadeh et al. 1998, J Clin Endocrinol Metab 83: 1911-1915) no statistically significant reduction in cholesterol has been shown. In animal models of obesity and obese diabetes, reductions in both total cholesterol and circulating triglycerides have been reported (Alemzadeh et al. European Journal of Endocrinology (2002) 146 871-879; Alemzadeh and Tushaus Med Sci Monit, 2005; 11(12): BR439-444). In these models, diazoxide was administered when the treated animals transitioned from a normal phenotype to either obese or obese diabetic disease states.

Encapsulation system: A structural feature that contains drug within such as a pharmaceutical capsule. A gel into which drug is incorporated also is considered an encapsulation system.

Endogenous hyperlipidemia: characterized primarily by elevated VLDL (both triglycerides and cholesterol associated with VLDL may also be elevated). Endogenous hyperlipidemia results from an endogenous metabolic source of the increased lipids rather than from a dietary source of the increased lipids.

Equivalent amount: An amount of a derivative of a drug that in assays or upon administration to a subject produces an equal effect to a defined amount of the non-derivatized drug.

Fatty acid synthase: The central enzyme of a multienzyme complex that catalyses the formation of palmitate from acetylcoenzyme A, malonylcoenzyme A, and NADPH.

Fibrates: A class of amphipathic carboxylic acids primarily used for decreasing serum triglycerides, while increasing high density lipoprotein (HDL).

Fructose-1,6-bisphosphatase inhibitor: A class of anti-diabetic drugs which act by inhibiting fructose-1,6-bisphosphatase, a key enzyme in the gluconeogenesis pathway. Inhibition of this enzyme lowers elevated glucose levels typical of type II diabetes.

Gastric Lipase: An enzyme secreted into the gastrointestinal tract that catalyzes the hydrolysis of dietary triglycerides.

Glidant: An inactive component of a pharmaceutical formulation that prevents caking of the matrix during processing steps.

GLP-1: Glucagon like peptide 1 is a peptide produced by intestinal endocrine cells in two principle forms, GLP-1(7-36 amide) and GLP-1(7-37), upon the ingestion of food. It is involved in stimulation of glucose-dependent insulin secretion and insulin biosynthesis, inhibition of glucagon secretion and gastric emptying, and inhibition of food intake.

HDL cholesterol: The cholesterol found in high density lipoprotein particles. HDL particles are believed to transport cholesterol from tissues in the body to the liver.

HMG-CoA reductase inhibitors (or statins) form a class of hypolipidemic agents, used as pharmaceutical agents to lower cholesterol levels in people with or at risk of cardiovascular disease. They lower cholesterol by inhibiting the enzyme HMG-CoA reductase, which is the rate-limiting enzyme of the mevalonate pathway of cholesterol synthesis.

Hypercholesterolemia: A condition characterized by elevated cholesterol.

Hyperinsulinemia: Excessively high blood insulin levels, which is differentiated from hyperinsulinism, excessive secretion of insulin by the pancreatic islets. Hyperinsulinemia may be the result of a variety of conditions, such as obesity and pregnancy.

Hyperinsulinism: Excessive secretion of insulin by the pancreatic islets.

Hyperlipidemia: A general term for elevated concentrations of any or all of the lipids in the plasma, such as cholesterol, triglycerides and lipoproteins.

Hyperphagia: Ingestion of a greater than optimal quantity of food.

Hypertension: Chronic high blood pressure, typically above 140 (systolic) over 90 (diastolic) mm Hg. Chronically elevated blood pressure can cause blood vessel changes in the back of the eye, thickening of the heart muscle, kidney failure, and brain damage.

Hypertriglyceridemia (or hypertriglyceridaemia) denotes high (hyper-) blood levels (-emia) of triglycerides without elevated cholesterol, the most abundant fatty molecule in most organisms. It has been associated with atherosclerosis, even in the absence of hypercholesterolemia (high cholesterol levels). It can also lead to pancreatitis in excessive concentrations.

Hypoglycemia-associated autonomic failure: A condition where hypoglycemic patients, e.g., patients with type I diabetes or advanced type II diabetes, become unable to recognize the symptoms of hypoglycemia, and/or lose counter-regulatory hormonal responses, primarily glucagon and epiepherine extending periods of hypoglycemia with their associated sequelae.

Ingredient of a pharmaceutical composition: Refers to one or more materials used in the manufacture of a pharmaceutical composition. Ingredient can refer to an active ingredient (an agent) or to other materials in the compositions. Ingredients can include water and other solvents, salts, buffers, surfactants, non-aqueous solvents, and flavorings.

Insulin dosing: The administration of exogenous insulin to a subject.

Insulin resistance: A condition in which the tissues of the body are diminished in their ability to respond to insulin.

Ischemic injury: Injury to tissue that results from a low oxygen state usually due to obstruction of the arterial blood supply or inadequate blood flow leading to hypoxia in the tissue.

Ketoacidosis: Acidosis accompanied by the accumulation of ketone bodies (ketosis) in the body tissue and fluids, as in diabetic acidosis.

Kit: Refers to a packaged combination. A packaged combination can optionally include a label or labels, instructions and/or reagents for use with the combination.

Kir: Pore forming subunit of the KATP channel. Also known as the inwardly rectifying subunit of the KATP channel. Typically existing as Kir6.x and infrequently as Kir2.x subspecies.

KATP channel: An ATP sensitive potassium ion channel across the cell membrane formed by the association of 4 copies of a sulfonylurea receptor and 4 copies of a pore forming subunit Kir. Agonizing the channel can lead to membrane hyperpolarization.

KATP channel opener: As used herein refers to a compound of any of Formulae I-VIII, or a derivative, salt or prodrug thereof, having one or more or preferably all of the following three properties: (1) opening SURx/Kir6.y potassium channels, where x=1, 2A or 2B and y=1 or 2; (2) binding to the SURx subunit of KATP channels; and (3) inhibiting glucose induced release of insulin following administration of the compound in vivo.

Late myocardial remodeling: myocardial infarction induced alterations of left ventricular (LV) architecture with scar formation, ventricular dilatation, and hypertrophy of the noninfarcted (remote) myocardium.

Leptin: Product (16 kD) of the ob (obesity) locus. It is found in plasma of mammals and exerts a hormonal action, which reduces food uptake and increases energy expenditure.

Lipogenesis: The generation of new lipids, primarily triacylglycerides. It is dependent on the action of multiple distinct enzymes and transport molecules.

Lipolysis: The breakdown of fat by the coordinated action of multiple enzymes.

Lipoprotein lipase: An enzyme of the hydrolase class that catalyses the reaction of triacyglycerol and water to yield diacylglycerol and a fatty acid anion. The enzyme hydrolyses triacylglycerols in chylomicrons, very-low-density lipoproteins, low-density lipoproteins, and diacylglycerols.

Lubricant: An inactive component of a pharmaceutical formulation that provides for the flow of materials in various processing steps, particularly tableting.

Meglitinide: A class of anti-diabetic drugs which block potassium channels by antagonism of sulfonylurea receptor component of the channel in beta cells, resulting in increased insulin secretion.

Microparticle: A small particulate formed in the process of developing pharmaceutical formulations that may be coated prior to producing the final dosage from.

MTP (microsomal triglyceride transfer protein) inhibitors: A class of pharmaceutical compounds that inhibit MTP activity to lower cholesterol and triglycerides.

Nonalcoholic steatohepatitis (NASH): Nonalcoholic steatohepatitis or NASH is a common, silent liver disease. It resembles alcoholic liver disease, but occurs in people who drink little or no alcohol. The major features in NASH are fat in the liver, inflammation and liver damage. The disease may progress to cirrhosis.

Obesity: An increase in body weight beyond the limitation of skeletal and physical requirement, as the result of an excessive accumulation of fat in the body. Formally defined as having a body mass index greater than 30 kg/m2.

Obesity Prone: Subjects who because of genetic predisposition or prior history of obesity are at above average risk of becoming obese.

Obesity related co-morbidities: Any disease or condition of animals or humans that are increased incidence in obese or overweight subjects. Examples of such conditions include hypertension, prediabetes, type 2 diabetes, osteoarthritis and cardiovascular conditions.

Oral anti-hypertensive agent: An orally administered therapeutic agent which is used to treat high blood pressure. Oral anti-hypertensive agents suitable for use in methods described herein include but are not limited to diuretics, beta-blockers, calcium channel blockers, ACE inhibitors, rennin inhibitors, angiotensin II antagonists, and alpha-1 blockers.

Osmotically controlled release: A pharmaceutical dosage form in which the release of the active drug is principally achieved by the hydration of a swellable component of the formulation.

Overweight: An subject whose weight is above that which is ideal for their height but who fails to meet the criteria for classification as obese. In humans using Body Mass Index (kg/m2) an overweight subjects has a BMI between 25 and 30.

Oxidation of Fat: A series of reactions involving acyl-coenzyme A compounds, whereby these undergo beta oxidation and thioclastic cleavage, with the formation of acetyl-coenzyme A; the major pathway of fatty acid catabolism in living tissue.

Pharmaceutical composition: Refers to a composition that contains an agent and one or more other ingredients that is formulated for administration to a subject. An agent refers to an active ingredient of a pharmaceutical composition. Typically active ingredients are active for treatment of a disease or condition. For example, agents that can be included in pharmaceutical compositions include agents for treating obesity or diabetes. The pharmaceutically active agent can be referred to as “a pharmaceutical active.”

Pharmaceutical effect: Refers to an effect observed upon administration of an agent intended for treatment of a disease or disorder or for amelioration of the symptoms thereof.

Pharmacodynamic: An effect mediated by drug action.

Pharmacokinetic: Relating to the absorption, distribution, metabolism and elimination of the drug in the body.

Poly-cystic ovarian syndrome: A disorder where the ovary secretes abnormally high levels of testosterone and estrogens, which results in irregular or no menses, excess body hair growth, occasionally baldness, and often obesity, diabetes and hypertension.

PEO: An abbreviation for polyoxyethylene. As used herein, and except as otherwise indicated, polyoxyethylene is a polyether polymer of ethylene glycol having an average molecular weight of greater than 20,000 g/mol. In some embodiments, the average molecular weight of PEO is from greater than 20,000 up to 300,000 g/mol. PEO may be used in the form of copolymers with other polymers or as a mixture of a combination of grades. Preferred concentrations of PEO include 3% to 40% either as a single grade or as a mixture of grades (molecular weights).

Polymorph: A crystalline form of a compound that exists in at least two crystalline forms. Polymorphic forms of any given compound are defined by the same chemical formula and/or composition and are as distinct in chemical structure as crystalline structures of two different chemical compounds. Such compounds may differ in packing or geometrical arrangement of respective crystalline lattices. The chemical and/or physical properties or characteristics of the various polymorphs may vary with each distinct polymorphic form, and may include, but are not limited to, variations in solubility, melting point, density, hardness, crystal shape, optical and electrical properties, vapor pressure, and stability.

Preadipocyte: A progenitor cell to adipocytes.

PPAR agonist: Peroxisome proliferator-activated receptor agonists include compounds that activate, e.g. the PPARγ receptor, leading to, e.g., a decrease in insulin resistance, modification of adipocyte differentiation, and a decrease in leptin levels. Thiazolidinediones target PPARγ.

PPAR partial agonist: Peroxisome proliferator-activated receptor partial agonists are PPAR agonists that can only incompletely agonize the receptor.

Prediabetic: A condition that precedes diagnosis of type II diabetes. Type II diabetes is a form of diabetes mellitus which is characterized by insulin insensitivity or resistance.

Prodrug: Refers to a compound which, when metabolized, yields the desired active compound. Typically, the prodrug is inactive, or less active than the active compound, but may provide advantageous handling, administration, or metabolic properties. For example, some prodrugs are esters of the active compound; during metabolysis, the ester group is cleaved to yield the active drug. Also, some prodrugs are activated enzymatically to yield the active compound, or a compound which, upon further chemical reaction, yields the active compound.

Prolonged Administration (prolonged basis): Administration of a pharmaceutically acceptable formulation of a drug for 7 or more days. Typically, prolonged administration is for at least two weeks, preferably at least one month, and even more preferably at least two months (i.e. at least 8 weeks).

Quick dissolving formulation: A pharmaceutical formulation which upon oral administration may release substantially all of the drug active from the formulation within 10 minutes.

QD, BID and TID: QD (quaque die) means once per day administration of a pharmaceutical substance or formulation; BID (bis in die) means twice per day; and TID (Ter In Die) means three time per day.

Release formulation (sustained), (or “sustained release formulation”): A formulation of pharmaceutical product that, upon administration to animals, provides for release of the active pharmaceutical over an extended period of time than provided by formulations of the same pharmaceutical active that result in rapid uptake. Similar terms are extended-release, prolonged-release, and slow-release. In all cases, the preparation, by definition, has a reduced rate of release of active substance.

Release formulation (delayed), (or “delayed release formulation”): Delayed-release products are modified-release, but are not extended-release. They involve the release of discrete amount(s) of drug some time after drug administration, e.g. enteric-coated products, and exhibit a lag time during which little or no absorption occurs.

Release formulation (controlled), (or “controlled release formulation”): A formulation of pharmaceutical product that may include both delay of release of pharmaceutical active upon administration and control of release in the manner described for sustained release.

Salt: The neutral, basic or acid compound formed by the union of an acid or an acid radical and a base or basic radical. Used generally to describe any ionic compound not containing an oxide or hydroxide ion.

A “short-acting anti-diabetic drug” refers to an agent that helps the pancreas produce insulin. Typically, a short-acting anti-diabetic drug is taken with meals to boost the insulin response to each meal. Short acting anti-diabetic drugs typically have rapid onset of effects with maximal effects typically realized between 30 minutes and 3 hours after administration and with circulating half-lives less than 5 hours.

Solid oral dosage form: Pharmaceutical formulations designed for oral administration including capsules and tablets.

Squalene synthase: squalene synthase is an enzyme at a branch point in the isoprenoid biosynthetic pathway capable of diverting carbon flow to the biosynthesis of sterols.

Squalene Synthase inhibitor: an inhibitor of squalene synthase.

Subject: Refers to animals, including mammals, such as human beings, domesticated animals, and animals of commercial value.

Sulfonylurea receptor: A component of the KATP channel responsible for interaction with sulfonylurea, other KATP channel antagonists, diazoxide and other KATP channel agonists.

Sulfonylurea receptor antagonist: a molecule which upon interaction with the sulfonylurea receptor acts to block the KATP channel.

Tablet: Pharmaceutical dosage form that is produced by forming a volume of a matrix containing pharmaceutical active and excipients into a size and shape suitable for oral administration.

Thermogenesis: The physiological process of heat production in the body.

Thiazolidinedione: A class of anti-diabetic drugs comprising a thiazolidine ring having 2 oxo groups attached. Thiazolidinediones bind to and activate the PPARγ receptor.

Threshold Concentration: The minimum circulating concentration of a drug required to exert a specific metabolic, physiological or compositional change in the body of a treated human or animal.

Thyroid receptor: The thyroid (hormone) receptor regulates a diverse set of genes that control processes from embryonic development to adult homeostasis. Receptors for thyroid hormones are members of a large family of nuclear receptors that include those of the steroid hormones. Upon binding of thyroid hormone, the thyroid receptor releases corepressor proteins and undergoes a conformational change that allows for the interaction of coactivating proteins necessary for gene transcription.

Thyroid receptor activator: A molecule which upon interaction with the thyroid hormone receptor acts to increase activity of the thyroid hormone receptor.

Treatment: Any manner in which the symptoms of a condition, disorder or disease or other indication, are ameliorated or otherwise beneficially altered.

Triglyceride: Storage fats of animal and human adipose tissue principally consisting of glycerol esters of saturated fatty acids.

Type I diabetes: A chronic condition in which the pancreas makes little or no insulin because the beta cells have been destroyed.

Type II diabetes: A chronic condition that is primarily characterized by insulin resistance, relative insulin deficiency, and hyperglycemia.

Uncoupling protein: A family of proteins that allow oxidation in mitochondria to proceed without the usual concomitant phosphorylation to produce ATP.

Visceral fat: Human adipose tissues principally found below the subcutaneous fat and muscle layer in the body.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows UV spectra of the free form diazoxide and the sodium and potassium salts of diazoxide in acetonitrile.

FIG. 2 shows UV spectra of the free form diazoxide at varying pH.

FIG. 3 shows UV spectra of the free form diazoxide and sodium and potassium salts of diazoxide in methanol.

FIG. 4A, FIG. 4B, FIG. 4C and FIG. 4D show X-Ray Powder Diffraction patterns for (a) free form diazoxide, (b) potassium salt of diazoxide from THF, (c) lysine salt of diazoxide from MEK, and (d) sodium salt of diazoxide from acetonitrile, respectively.

FIG. 5A, FIG. 5B and FIG. 5C show NMR spectra (DMSO-d6 solvent) for (a) free form diazoxide, (b) potassium salt, and (c) sodium salt, respectively.

FIG. 6A, FIG. 6B and FIG. 6C show X-Ray Powder Diffraction patterns for (a) sodium salt of diazoxide, (b) sodium salt of diazoxide after slurrying in water, and (c) free form diazoxide, respectively.

FIG. 7 shows DSC spectra for the free form diazoxide (top) and potassium salt of diazoxide (bottom). Description: “a” (Integral=−317.56 mJ; normalized=−84.82 Jg 1; Onset=120.81° C.; Peak=121.29° C.); “b” (Integral=−1170.43 mJ; normalized=154.64 Jg-1; Onset=329.54° C.; Peak=329.21° C.); “c” (Extrap. Peak=355.01° C.; Peak Value=−4.58 mW; normalized=−1.22 Wg-1; Peak=353.53° C.).

FIG. 8 shows TGA spectra for the free form diazoxide (top) and potassium salt of diazoxide (bottom).

FIG. 9A, FIG. 9B and FIG. 9C show X-Ray Powder Diffraction patterns for (a) potassium salt of diazoxide, (b) potassium salt of diazoxide after slurrying in toluene, and (c) potassium salt of diazoxide after slurrying in toluene for 14 days, respectively.

FIG. 10A, FIG. 10B and FIG. 10C show X-Ray Powder Diffraction patterns for (a) free form diazoxide, (b) choline salt of diazoxide, and (c) hexamethyl hexamethylene diammonium hydroxide salt of diazoxide, respectively.

FIG. 11 shows DSC spectra for the free form diazoxide (top) and choline salt of diazoxide (bottom). Description: “a” (Integral=−41.24 mJ; normalized=−8.05 Jg 1; Onset=101.23° C.; Peak=119.29° C.); “b” (Integral=−497.37 mJ; normalized=97.10 Jg-1; Onset=166.03° C.; Peak=167.27° C.); “c” (Integral=−1167.83 mJ; normalized=154.29 Jg-1; Onset=329.54° C.; Peak=329.21° C.).

FIG. 12 shows TGA spectra for the free form diazoxide (top) and choline salt of diazoxide (bottom).

FIG. 13A, FIG. 13B and FIG. 13C show X-Ray Powder Diffraction patterns for (a) choline salt of diazoxide, (b) choline salt of diazoxide after slurrying in dichloromethane for 7 days, and (c) choline salt of diazoxide after moisture sorption analysis, respectively.

FIG. 14A, FIG. 14B and FIG. 14C show NMR spectra (DMSO-d6 solvent) for (a) free form diazoxide, (b) choline salt, and (c) hexamethyl hexamethylene diammonium hydroxide salt of diazoxide, respectively.

FIG. 15A shows overlay XRPD patterns of free form diazoxide, the product of potassium methoxide in methanol, and the product of sodium methoxide in methanol. FIG. 15B, FIG. 15C and FIG. 15D show the XRPD patterns for product of potassium methoxide reaction with diazoxide in methanol, product of sodium methoxide reaction with diazoxide in methanol, and freeform diazoxide, respectively.

FIG. 16A and FIG. 16B show XRPD patterns of (a) polymorphic Form A of the choline salt of diazoxide, and (b) a mixture of polymorphic forms A and B of the choline salt of diazoxide, respectively.

FIG. 17A and FIG. 17B show the NMR spectra (DMSO-d6 solvent) for (a) polymorphic Form A of the choline salt of diazoxide, and (b) polymorphic Form B of the choline salt of diazoxide, respectively.

FIG. 18A, FIG. 18B and FIG. 18C show XRPD patterns of (a) polymorphic Form A of the potassium salt of diazoxide, (b) polymorphic Form B of the potassium salt of diazoxide, and (c) polymorphic Form C of the potassium salt of diazoxide, respectively.

FIG. 19A, FIG. 19B, FIG. 19C and FIG. 19D show XRPD patterns of (a) polymorphic Form D of the potassium salt of diazoxide, (b) polymorphic Form E of the potassium salt of diazoxide, (c) polymorphic Form F of the potassium salt of diazoxide, and (d) polymorphic Form G of the potassium salt of diazoxide, respectively.

FIG. 20 shows the DSC spectra of diazoxide choline salt Form A. Description: “a” (Extrap. Peak=120.44° C.; Peak Value=−1.02 mW; normalized=−0.20 Wg-1; Peak=118.63° C.); “b” (Extrap. Peak=167.94° C.; Peak Value=−19.39 mW; normalized=−3.79 Wg-1; Peak=167.27° C.).

FIG. 21 shows the DSC spectra of diazoxide choline salt Form B. Description: “a” (Extrap. Peak=165.05° C.; Peak Value=−3.85 mW; normalized=−0.86 Wg-1; Peak=162.66° C.).

FIG. 22 provides systolic blood pressure (SBP) and diastolic blood pressure (DBP) for Proglycem Oral Suspension (Proglycem) and Diazoxide Choline Controlled-Release Tablets (DCCRT) at various times following dose administration (mean±SEM).

FIG. 23 provides pulse rate for Proglycem Oral Suspension (Proglycem) and Diazoxide Choline Controlled-Release Tablets (DCCRT) at various times following dose administration (mean±SEM).

FIG. 24 provides mean plasma diazoxide (±SD) concentrations after a 200 mg dose of diazoxide (linear coordinates).

FIG. 25 provides mean plasma diazoxide (±SD) concentrations after a 200 mg dose of diazoxide (semilog coordinates).

FIG. 26 provides simulations to steady-state of once daily dosing with 200 mg diazoxide.

FIG. 27 provides a bar graph comparing the median percentage change in levels of different lipid measurements for patients that were treated with DCCR for 12 weeks followed by co-administration of DCCR with fenofibrate choline for 6 weeks, and patients that received placebo for 12 weeks followed by co-administration of placebo with fenofibrate choline for 6 weeks. Lipids measured for these studies were triglycerides (TG), total cholesterol (TC), non-HDL-C, VLDL-C, LDL-C, and HDL-C. Details are described in Example sections 15.1 and 15.2.

FIG. 28 provides a bar graph comparing the median percentage change in levels of different lipid measurements for patients that were treated with DCCR for 12 weeks followed by co-administration of DCCR with fenofibrate choline for 6 weeks, and patients that only received fenofibrate choline for 6 weeks. Lipids measured for these studies were triglycerides (TG), total cholesterol (TC), non-HDL-C, VLDL-C, LDL-C, and HDL-C. Details are described in Example section 15.1 and 15.3.

FIG. 29 provides a bar graph comparing the median percentage change in levels of different lipid measurements for patients that were treated with DCCR for 12 weeks followed by co-administration of DCCR with fenofibrate choline for 6 weeks, patients that received omega-3 fatty acids and fenofibrate for 8 weeks (FIG. 2 of Roth et al. J Cardiovasc Pharmacol. 2009 September; 54(3):196-203), and their respective controls. Lipids measured in these studies were triglycerides (TG), total cholesterol (TC), non-HDL-C, VLDL-C, LDL-C, and HDL-C. JCP=Data from Roth et al. (J Cardiovasc Pharmacol. 2009 September; 54(3):196-203). Details are described in section 15.1 and 15.4.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides salts of compounds of Formulae I-VIII and methods for their preparation. Salts of compounds of Formulae I-IV may be prepared using monovalent alkali metal cations and compounds which include one or more of a tertiary amine or quaternary ammonium moiety. In such salts, the compounds of Formulae I-IV exist in their anionic form. Furthermore, it has been discovered that the selection of a solvent for the preparation of these salts plays an important role in salt formation. Also described herein is the failure to obtain a salt of diazoxide from an alkali metal alkoxide using the method described in U.S. Pat. No. 2,986,573.

Compounds of Formulae V-VIII can form both anions and cations, and thus salts can be prepared using a variety of counter ions, including both anions and cations. Cations of the compounds of Formulae V-VIII can be formed at an amino group, and anions of the compounds of Formulae V-VIII can be formed at either an amino group or at the sulfonyl group. The formation of salts based on compounds of Formulae V-VIII can be done in a variety of solvents, preferably organic solvents.

As discussed herein, two polymorphic forms (i.e., Forms A and B) of the choline salt of diazoxide have been identified. In summary, both Forms A and B are anhydrous crystals of diazoxide choline salt. Diazoxide choline salt Form A can be formed using fast cooling procedures as provided herein, whereas slow cooling procedures generally favor formation of Form B. Slurry studies shows that Form A readily converts to Form B. Without wishing to be bound by theory, the slurry studies indicate that Form B of diazoxide choline salt is the thermodynamically more stable form.

Regarding the potassium salt of diazoxide, seven polymorphic forms have been identified (i.e., Forms A-G). Diazoxide potassium salt Forms C, D, and F were observed be an acetone solvent, a hemihydrate, and a dioxane solvent, respectively. Forms A, B, E, and G were not commonly observed during screening, and elemental analysis suggests that Forms A, B, E and G may be mixtures, have residual solvent present, and/or not be a potassium salt, at least in part. Without wishing to be bound by theory, slurry studies suggest that Form D is the thermodynamically most stable polymorph of the diazoxide potassium salt polymorphs.

Further provided are pharmaceutical formulations of particular KATP channel openers of salts of compounds of Formulae I-VIII that when administered to subjects achieve novel pharmacodynamic, pharmacokinetic, therapeutic, physiological, and metabolic outcomes. Yet further provided are pharmaceutical formulations, methods of administration and dosing of particular KATP channel openers selected from salts of the compounds defined by Formulae I-VIII that achieve therapeutic outcomes while reducing the incidence of adverse effects.

Still further provided are pharmaceutical formulations of particular KATP channel openers of salts of compounds of Formulae I-VIII that may be administered to subjects once per day (QD), twice per day (BID) or three time per day (TID). The skilled artisan would realize each dosage per day per time may comprise one or more tablets administered to a subject. In preferred embodiments, the total effective dose per day can be 100, 200, 300, 400, 500 or 600 mg of KATP channel openers as active agent (administered as control release formulations). Such doses can be effective in achieving weight loss or reduction in total cholesterol, reduction in LDL cholesterol, reduction in non-HDL cholesterol, reduction in VLDL cholesterol, increase in HDL cholesterol, and reduction in triglyceride without substantial caloric reduction, more preferably with no caloric reduction. Such control release formulations comprise particular KATP channel openers of salts of compounds of formulae I-VIII formulation that require once per day dosing with or without caloric restricted diet. In addition, with control release formulations, the dosing required per day to reach clinical efficacy is much less compared to immediate release formulations.

In particular, pharmaceutical formulations selected from salts of compounds defined by Formulae I-VIII and formulated for oral administration exhibit advantageous properties including: facilitating consistency of absorption, pharmacokinetic and pharmacodynamic responses across treated patients, contributing to patient compliance and improving the safety profile of the product, such as by reducing the frequency of serious adverse effects. Method of treatment of metabolic and other diseases of humans and animals by administering the formulations are also provided.

As shown below, diazoxide and derivatives thereof can exist as proton tautomers. Proton tautomers are isomers that differ from each other only in the location of a hydrogen atom and a double bond. The hydrogen atom and double bond switch locations between a carbon atom and a heteroatom, such as for example N. Thus, when the substituent on the nitrogen is hydrogen, the two isomeric chemical structures may be used interchangeably.

The particular KATP channel openers that can be used in the invention formulations include salts of any of the compounds within Formulae Ito VIII. Exemplary compounds which have been previously reported include diazoxide, BPDZ 62, BPDZ 73, NN414 and BPDZ 154 (see, for example, Schou et al., Bioorg. Med. Chem., 13, 141-155 (2005)). Compound BPDZ 154 also is an effective KATP channel activator in patients with hyperinsulinism and in patients with pancreatic insulinoma. The synthesis of BPDZ compound is provided in Cosgrove et al., J. Clin. Endocrinol. Metab., 87, 4860-4868 (2002).

Channel openers demonstrating decreased activity in the inhibition of insulin release and increased activity in vascular smooth muscle tissue have been previously reported and include analogs of diazoxide such as, for example, 3-isopropylamino-7-methoxy-4H-1,2,4,-benzothiadiazine 1,1-dioxide, (a selective Kir6.2/SUR1 channel opener; see Dabrowski et al., Diabetes, 51, 1896-1906 (2002), and 2-alkyl substituted diazoxides (see, for example, Ouedraogo et al., Biol. Chem., 383, 1759-1768 (2002)). The 2-alkyl substituted diazoxides generally do not function as traditional potassium channel activators, but instead show potential as Ca2+ blockers.

Other diazoxide analogs which have been previously reported include described in Schou et al., Bioorg. Med. Chem., 13, 141-155 (2005), are shown below.

-   -   R¹, R² and R³ are:     -   a) H, Cl, NHCH(CH₃)₂     -   b) CF₃, H, NHCH(CH₃)₂     -   c) H, Cl, NHCH₂CH₂CH(CH₃)₂     -   d) H, Cl, NH-cyclobutyl

Diazoxide analogs having different alkyl substituents at the 3 position of the molecule (identified as R³ shown below) are described in Bertolino et al., Receptors and Channels, 1, 267-278 (1993).

-   -   R³, R⁶ and R⁷ are:     -   a) H, H, CH₃     -   b) H, H, Cl     -   c) CH₃, Cl, H     -   d) CH₂Cl, H, Cl     -   e) NH₂, H, H     -   f) CH₂CH₂Cl, H, Cl     -   g) nC₄H₉, H, Cl     -   h) nC₅H₁₁, H, Cl     -   i) nC₇H₁₅, H, Cl     -   j) nC₃H₇, Cl, H     -   k) nC₄H₉, Cl, H     -   l) nC₅H₁₁, Cl, H     -   m) nC₇H₁₅, Cl, H     -   n) nC₃H₇, Cl, Cl     -   o) nC₄H₉, Cl, Cl     -   p) nC₅H₁₁, Cl, Cl     -   q) nC₇H₁₅, Cl, Cl     -   r) H, CL, H

KATP channel activity of salts of the compounds of Formulae I-VIII and related compounds can be measured by membrane potential studies as described in Schou et al., Bioorg. Med. Chem., 13, 141-155 (2005) and Dabrowski, et al., Diabetes, 51, 1896-1906 (2002).

Measurement of the inhibition of glucose-stimulated insulin release from βTC6 cells is described in Schou et al., Bioorg. Med. Chem., 13, 141-155 (2005). The ability of particular KATP channel openers to inhibit release of insulin from incubated rat pancreatic islets can be performed as described by Ouedraogo et al., Biol. Chem., 383, 1759-1768 (2002).

Activation of recombinant KATP channels by KATP channel openers can be examined by monitoring macroscopic currents of inside-out membrane patches from Xenopus oocytes co-expressing Kir6.2 and either SUR1, SUR2A or SUR2B. SUR expressing membranes can be prepared by known methods. See, for example, Dabrowski et al., Diabetes, 51, 1896-1906 (2002).

Binding experiments can be used to determine the ability of KATP channel openers to bind SUR1, SUR2A and SUR2B. See, for example, Schwanstecher et al., EMBO J., 17, 5529-5535 (1998).

Preparation of SUR1 and SUR2A chimeras, as described by Babenko et al., allows for comparison of pharmacologic profiles (i.e. sulfonyl sensitivity and responsiveness to diazoxide or other potassium channel openers) of the SUR1/Kir6.2 and SUR2A/Kir6.2 potassium channels. See Babenko et al., J. Biol. Chem., 275(2), 717-720 (2000). The cloning of a sulfonylurea receptor and an inwardly rectifying K⁺ channel is described by Isomoto et al., J. Biol. Chem., 271 (40), 24321-24324 (1996); D'hahan et al., PNAS, 96(21), 12162-12167 (1999).

Differences between the human SUR1 and human SUR2 genes are described and shown in Aguilar-Bryan et al., Physiological Review, 78(1):227-245 (1998).

“Halo” and “halogen” refer to all halogens, that is, chloro (Cl), fluoro (F), bromo (Br), or iodo (I).

“Hydroxyl” and “hydroxy” refer to the group —OH.

“Substituted oxy” refers to the group —ORaa, where Raa can be alkyl, substituted alkyl, acyl, substituted acyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, aralkyl, substituted aralkyl, cycloalkyl, substituted cycloalkyl, heterocyclyl, or substituted heterocyclyl.

“Substituted thiol” refers to the group-SRbb, where Rbb can be alkyl, substituted alkyl, acyl, substituted acyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, aralkyl, substituted aralkyl, cycloalkyl, substituted cycloalkyl, heterocyclyl, or substituted heterocyclyl.

“Alkyl” refers to an alkane-derived radical containing from 1 to 10, preferably 1 to 6, more preferably 1-4, yet more preferably 1-2, carbon atoms. Alkyl includes straight chain alkyl, branched alkyl and cycloalkyl, such as methyl, ethyl, propyl, isopropyl, butyl, t-butyl, and the like. The alkyl group can be attached at any available point to produce a stable compound. An “alkylene” is a divalent alkyl.

A “substituted alkyl” is an alkyl group independently substituted with 1 or more, e.g., 1, 2, or 3, groups or substituents such as halo, hydroxy, optionally substituted alkoxy, optionally substituted alkylthio, alkylsulfinyl, alkylsulfonyl, optionally substituted amino, optionally substituted amido, amidino, urea optionally substituted with alkyl, aminosulfonyl optionally N-mono- or N,N-di-substituted with alkyl, alkylsulfonylamino, carboxyl, heterocycle, substituted heterocycle, nitro, cyano, thiol, sulfonylamino or the like attached at any available point to produce a stable compound. In particular, “fluoro substituted” refers to substitution by 1 or more, e.g., 1, 2, or 3 fluorine atoms. “Optionally fluoro substituted” means that substitution, if present, is fluoro. The term “optionally substituted” as used herein means that substitution may, but need not, be present.

“Lower alkyl” refers to an alkyl group having 1-6 carbon atoms.

A “substituted lower alkyl” is a lower alkyl which is substituted with 1 or more, e.g., 1, 2, or 3, groups or substituents, as defined above, attached at any available point to produce a stable compound.

“Cycloalkyl” refers to saturated or unsaturated, non-aromatic monocyclic, bicyclic or tricyclic carbon ring systems of 3-8, more preferably 3-6, ring members per ring, such as cyclopropyl, cyclopentyl, cyclohexyl, adamantyl, and the like. “Cycloalkylene” is a divalent cycloalkyl.

“Substituted cycloalkyl” refers to saturated or unsaturated, non-aromatic monocyclic, bicyclic or tricyclic carbon ring systems of 3-8, more preferably 3-6, ring members per ring, such as cyclopropyl, cyclopentyl, cyclohexyl, adamantyl, and the like independently substituted with 1 or more, e.g., 1, 2, or 3, groups or substituents such as halo, hydroxy, optionally substituted alkoxy, optionally substituted alkylthio, alkylsulfinyl, alkylsulfonyl, optionally substituted amino, optionally substituted amido, amidino, urea optionally substituted with alkyl, aminosulfonyl optionally N-mono- or N,N-di-substituted with alkyl, alkylsulfonylamino, carboxyl, heterocycle, substituted heterocycle, nitro, cyano, thiol, sulfonylamino or the like attached at any available point to produce a stable compound.

“Aryl” alone or in combination means phenyl or naphthyl optionally carbocyclic fused with a cycloalkyl of preferably 5-7, more preferably 5-6, ring members.

“Substituted aryl” refers to an aryl group as defined above independently substituted with 1 or more, e.g., 1, 2, or 3, groups or substituents such as halo, hydroxy, optionally substituted alkoxy, optionally substituted alkylthio, alkylsulfinyl, alkylsulfonyl, optionally substituted amino, optionally substituted amido, amidino, urea optionally substituted with alkyl, aminosulfonyl optionally N-mono- or N,N-di-substituted with alkyl, alkylsulfonylamino, carboxyl, heterocycle, substituted heterocycle, nitro, cyano, thiol, sulfonylamino or the like attached at any available point to produce a stable compound.

“Alkoxy” denotes the group —ORcc, where Rcc is alkyl. “Lower alkoxy” denotes the group —ORccc, where Rccc is lower alkyl

“Substituted alkoxy” denotes the group —ORdd, where Rdd is substituted alkyl. “Substituted lower alkoxy” denotes the group —ORddd, where Rddd is substituted lower alkyl.

“Alkylthio” or “thioalkoxy” refers to the group —S—Ree, where Ree is alkyl.

“Substituted alkylthio” or “substituted thioalkoxy” refers to the group —S—R, where R is substituted alkyl.

“Sulfinyl” denotes the group —S(O)—.

“Sulfonyl” denotes the group —S(O)₂—.

“Substituted sulfinyl” denotes the group —S(O)—Rff, where Rff is alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, cycloalkylalkyl, substituted cycloalkylalkyl, heterocyclyl, substituted heterocyclyl, heterocyclylalkyl, substituted hetereocyclylalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heteroaralkyl, substituted heteroaralkyl, aralkyl or substituted aralkyl.

“Substituted sulfonyl” denotes the group —S(O)₂Rgg, where Rgg is alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, cycloalkylalkyl, substituted cycloalkylalkyl, heterocyclyl, substituted heterocyclyl, heterocyclylalkyl, substituted hetereocyclylalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heteroaralkyl, substituted heteroaralkyl, aralkyl or substituted aralkyl.

“Sulfonylamino” denotes the group —S(O)₂NRhh- where Rhh is hydrogen or alkyl.

“Substituted sulfonylamino” denotes the group —S(O)2NRii-Rjj, where Rii is hydrogen or optionally substituted alkyl, and Rjj is alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocyclyl, substituted heterocyclyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heteroaralkyl, substituted heteroaralkyl, aralkyl or substituted aralkyl.

“Amino” or “amine” denotes the group —NH2. A “divalent amine” denotes the group —NH—. A “substituted divalent amine” denotes the group —NRkk- wherein Rkk is alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, acyl, substituted acyl, sulfonyl or substituted sulfonyl.

“Substituted amino” or “substituted amine” denotes the group —NRmmRnn, wherein Rmm and Rnn are independently hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, acyl, substituted acyl, sulfonyl, substituted sulfonyl, or cycloalkyl provided, however, that at least one of Rmm and Rnn is not hydrogen. RmmRnn in combination with the nitrogen may form an optionally substituted heterocyclic or heteroaryl ring.

“Alkylsulfinyl” denotes the group —S(O)Roo, wherein Roo is optionally substituted alkyl.

“Alkylsulfonyl” denotes the group —S(O)2Rpp, wherein Rpp is optionally substituted alkyl.

“Alkylsulfonylamino” denotes the group —NRqqS(O)2Rrr, wherein Rrr is optionally substituted alkyl, and Rqq is hydrogen or alkyl.

A “primary amino substituent” denotes the group —NH2.

A “secondary amino substituent” denotes the group —NHRss, wherein Rss is alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, acyl, substituted acyl, sulfonyl, substituted sulfonyl, or cycloalkyl.

A “tertiary amino substituent” denotes the group —NRssRtt, wherein Rss and Rtt are independently alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, acyl, substituted acyl, sulfonyl, substituted sulfonyl, or cycloalkyl.

“Quaternary ammonium substituent” denotes the group —N+RssRttRuu, wherein Rss, Rtt and Ruu are independently alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, acyl, substituted acyl, sulfonyl, substituted sulfonyl, or cycloalkyl.

“Heteroaryl” means a monocyclic aromatic ring structure containing 5 or 6 ring atoms, or a bicyclic aromatic group having 8 to 10 atoms, containing one or more, preferably 1-4, more preferably 1-3, even more preferably 1-2, heteroatoms independently selected from the group consisting of O, S, and N. Heteroaryl is also intended to include oxidized S or N, such as sulfinyl, sulfonyl and N-oxide of a tertiary ring nitrogen. A carbon or nitrogen atom is the point of attachment of the heteroaryl ring structure such that a stable aromatic ring is retained. Examples of heteroaryl groups are pyridinyl, pyridazinyl, pyrazinyl, quinaoxalyl, indolizinyl, benzo[b]thienyl, quinazolinyl, purinyl, indolyl, quinolinyl, pyrimidinyl, pyrrolyl, oxazolyl, thiazolyl, thienyl, isoxazolyl, oxathiadiazolyl, isothiazolyl, tetrazolyl, imidazolyl, triazinyl, furanyl, benzofuryl, indolyl, and the like. “Heteroarylene” means a divalent heteroaryl.

“Heterocycle” or “heterocyclyl” means a saturated or unsaturated, non-aromatic carbocyclic group having a single ring or multiple condensed rings, e.g. a cycloalkyl group having from 5 to 10 atoms in which from 1 to 3 carbon atoms in a ring are replaced by heteroatoms, such as O, S, N, and are optionally fused with benzo or heteroaryl of 5-6 ring members and/or are optionally substituted. Heterocyclyl is intended to include oxidized S or N, such as sulfinyl, sulfonyl and N-oxide of a tertiary ring nitrogen. Examples of heterocycle or heterocyclyl groups are morpholino, tetrahydrofuranyl, dihydropyridinyl, piperidinyl, pyrrolidinyl, piperazinyl, dihydrobenzofuryl, dihydroindolyl, and the like.

“Heterocyclylalkyl” refers to the group —R-Het where Het is a heterocycle group and R is an alkylene group.

A “substituted heteroaryl,” “substituted heterocyclyl,” or “substituted heterocyclylalkyl” is a heteroaryl, heterocyclyl, or heterocyclylalkyl, respectively, independently substituted with 1 or more, e.g., 1, 2, or 3, groups or substituents such as halogen, hydroxy, optionally substituted alkoxy, optionally substituted alkylthio, alkylsulfinyl, alkylsulfonyl, acyloxy, optionally substituted aryl, optionally substituted aryloxy, optionally substituted heteroaryloxy, optionally substituted amino, optionally substituted amido, amidino, urea optionally substituted with alkyl, aryl, heteroaryl or heterocyclyl groups, aminosulfonyl optionally N-mono- or N,N-di-substituted with alkyl, aryl or heteroaryl groups, alkylsulfonylamino, arylsulfonylamino, heteroarylsulfonylamino, alkylcarbonylamino, arylcarbonylamino, heteroarylcarbonylamino, carboxyl, heterocycle, substituted heterocycle, heteroaryl, substituted heteroaryl, nitro, cyano, thiol, sulfonylamino, optionally substituted alkyl, optionally substituted alkenyl, or optionally substituted alkynyl, attached at any available point to produce a stable compound.

“Amido” denotes the group —C(O)NH2. “Substituted amido” denotes the group —C(O)NRkRl, wherein Rk and Rl are independently hydrogen, lower alkyl, substituted lower alkyl, aryl, substituted aryl, heteroaryl, or substituted heteroaryl, provided, however, that at least one of Rk and Rl is not hydrogen. RkRl in combination with the nitrogen may form an optionally substituted heterocyclic or heteroaryl ring.

“Amidino” denotes the group —C(═NRm)NRnRo, wherein Rm, Rn, and Ro are independently hydrogen or optionally substituted lower alkyl.

“Acyloxy” denotes the group —OC(O)Rh, where Rh is hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocyclyl, substituted heterocyclyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl and the like.

“Aryloxy” denotes the group —OAr, where Ar is an aryl, or substituted aryl, group. “Heteroaryloxy” denotes groups —OHet, wherein Het is an optionally substituted heteroaryl group.

“Arylsulfonylamino” denotes the group —NRqS(O)2Rs, wherein Rs is optionally substituted aryl, and Rq is hydrogen or lower alkyl.

“Heteroarylsulfonylamino” denotes the group —NRqS(O)2Rt, wherein Rt is optionally substituted heteroaryl, and Rq is hydrogen or lower alkyl.

“Alkylcarbonylamino” denotes the group —NRqC(O)Rp, wherein Rp is optionally substituted alkyl, and Rq is hydrogen or lower alkyl.

“Arylcarbonylamino” denotes the group —NRqC(O)Rs, wherein Rs is optionally substituted aryl, and Rq is hydrogen or lower alkyl.

“Heteroarylcarbonylamino” denotes the group —NRqC(O)Rt, wherein Rt is optionally substituted aryl, and Rq is hydrogen or lower alkyl.

Pharmaceutical formulations containing KATP channel openers can include the free base of a compound defined by any of Formulae I-VIII, or a salt thereof. Salts of the compounds of Formulae I-VIII as provided herein may have one or more of the following characteristics: (1) stability in solution during synthesis and formulation, (2) stability in a solid state, (3) compatibility with excipients used in the manufacture of tablet formulations, (4) quantitatively yield the KATP channel opener upon exposure to simulated or actual gastric and duodenal conditions, (5) release KATP channel opener from sufficiently small particles that are readily dissolved and absorbed, (6) provide, when incorporated into a pharmaceutical formulation, for absorption of greater than 80% of the administered dose, (7) present no elevated toxicological risk as compared to the free base of the KATP channel opener, (8) can be formulated into acceptable pharmaceutical formulations to treat obesity and other diseases of humans, (9) are acceptable to the FDA as the basis of a drug product, (10) can be recrystallized to improve purity, (11) can be used to form co-crystals of two or more salts of the KATP channel opener, (12) have limited hygroscopicity to improve stability, (13) synthetic and crystallization conditions under which the salt is formed can be varied resulting in different crystal structures (polymorphs) can be controlled in the synthesis of the salt, or (14) have improved solubility as compared to the free base in aqueous systems at physiological pH values.

The KATP channel openers provided in Formulae I-VIII are preferably formulated as pharmaceutically acceptable salts. Pharmaceutically acceptable salts are non-toxic salts in the amounts and concentrations at which they are administered. The preparation of such salts can facilitate the pharmacological use by altering the physical characteristics of a compound without preventing it from exerting its physiological effect. Useful alterations in physical properties include lowering the melting point to facilitate transmucosal administration and increasing the solubility to facilitate administering lower effective doses of the drug.

Salts of the compounds of Formulae I-IV can include metal cations, preferably alkali metal cations, such as for example, sodium or potassium. Cations can be selected from any group I alkali metal. Divalent metals cations, such as alkaline earth metals (e.g., magnesium, calcium and the like), have not been found to be useful for salt formation with the compounds of Formulae I-IV.

Salts of the compounds of Formulae I-IV which include alkali metal cations can be prepared by reacting the compounds of Formulae I-IV with an alkali metal hydroxide or alkali metal alkoxide, such as for example, NaOH, KOH or NaOCH3, in a variety of solvents which may be selected from low molecular weight ketones (e.g., acetone, methyl ethyl ketone), tetrahydrofuran (THF), dimethylformamide (DMF), and n-methylpyrrolidinone, and the like. Surprisingly, salt formation with an alkali metal hydroxide or alkoxide is not observed when an alcohol, particularly a lower alcohol such as for example methanol or ethanol, is used as the solvent. This result was confirmed by both X-Ray Powder Diffraction and NMR, and is contrary to the disclosure of U.S. Pat. No. 2,986,573, which purports to describe formation of diazoxide salts in alcohol.

The compounds of Formulae I-IV can also form salts with organic cations that include at least one tertiary amine or ammonium cation. Organic cation compounds can be monovalent, divalent, trivalent and tetravalent by inclusion of one, two, three or four tertiary amine or ammonium ions within the compound, respectively. When a multivalent compound is used, the tertiary amine or quaternary ammonium moieties are preferably separated by a chain of at least 4 atoms, more preferably by a chain of at least 6 atoms, such as for example, hexamethyl hexamethylene diammonium dihydroxide, wherein the quaternary ammonium moieties are separated by —(CH2)6—. Primary and secondary amines do not to effectively form salts with the compounds of Formulae I-IV.

Salts of the compounds of Formulae I-IV can be prepared by reacting the compounds of Formulae I-IV with compounds that include at least one tertiary amine or quaternary ammonium ion (e.g., choline hydroxide, hexamethylhexamethylene diammonium dihydroxide) in a solvent selected from low molecular weight ketones (e.g., acetone, methyl ethyl ketone), tetrahydrofuran, dimethylformamide, and n-methyl pyrrolidinone. As with the preparation of salts from alkali metal hydroxides, amine and ammonium containing compounds do not form salts when the solvent is an alcohol.

Pharmaceutically acceptable salts of the compounds of Formulae I-IV can also include basic addition salts such as those containing benzathine, chloroprocaine, choline, diethylamino-ethanol, hydroxyethyl pyrrolidine, ammonium, tetrapropylammonium, tetrabutylphosphonium, hexamethyl diammonium, methyldiethanamine, triethylamine, meglumine, and procaine, and can be prepared using the appropriate corresponding bases.

Preferred basic addition salts of the compounds of Formulae I-IV can include those containing hexamethyl hexamethylene diammonium, choline, sodium, potassium, methyldiethyl amine, triethylamine, diethylamino-ethanol, hydroxyethyl pyrrolidine, tetrapropylammonium and tetrabutylphosphonium ions.

Preferred basic addition salts of the compounds of Formulae I-IV can be prepared using hexamethyl hexamethylene diammonium dihydroxide, choline hydroxide, sodium hydroxide, sodium methoxide, potassium hydroxide, potassium methoxide, ammonium hydroxide, tetrapropylammonium hydroxide, and tetrabutylphosphonium hydroxide. The basic addition salts can be separated into inorganic salts (e.g., sodium, potassium and the like) and organic salts (e.g., choline, hexamethyl hexamethylene diammonium hydroxide, and the like).

The compounds of Formulae V-VIII have the unique property of being able to form both anions and cations. In basic media, the compounds of Formulae V-VIII typically form anions. Anions can be formed at either an amino or substituted amino substituent, or at the sulfonyl group. In acidic media, the compounds of Formulae V-VIII generally form cations by protonation of an amino group, thereby forming an ammonium moiety.

Salts of the anions of compounds of Formulae V-VIII can include metal cations, including monovalent metal cations of any group I alkali metal (e.g., sodium, potassium, and the like), divalent metal cations of any group II alkaline earth metal (e.g., calcium, magnesium, and the like), and aluminum cations.

Salts of the compounds of Formulae V-VIII which include metal cations can be prepared by reacting the compounds of Formulae V-VIII with a alkali or alkaline earth metal hydroxides or alkoxides, such as for example, sodium hydroxide or sodium methoxide, in an organic solvent, such as for example lower alcohols, low molecular weight ketones (e.g., acetone, methyl ethyl ketone, and the like), tetrahydrofuran, dimethylformamide, and n-methylpyrrolidinone, and the like.

Salts of the compounds of Formulae V-VIII, may include organic or inorganic counter ions, including but not limited to, acetate, acetonide, acetyl, adipate, aspartate, besylate, biacetate, bitartrate, bromide, butoxide, butyrate, calcium, camsylate, caproate, carbonate, citrate, cypionate, decanoate, diacetate, dimegulumine, dinitrate, dipotassium, dipropionate, disodium, disulfide, edisylate, enanthate, estolate, etabonate, ethylsuccinate, fumarate, furoate, gluceptate, gluconate, hexacetonide, hippurate, hyclate, hydrobromide, hydrochloride, isethionate, lactobionate, malate, maleate, meglumine, methylbromide, methylsulfate, metrizoate, nafate, napsylate, nitrate, oleate, palmitate, pamoate, phenpropionate, phosphate, pivalate, polistirex, polygalacturonate, probutate, propionate, saccharate, sodium glycinate, sodium phosphate, sodium succinate, stearate, succinate, sulfate, sulfonate, sulfosalicylate, tartrate, tebutate, terephalate, terephthalate, tosylate, triflutate, trihydrate, trisilicate, tromethamine, valerate, or xinafoate. Preferred organic cations include compounds having tertiary amines or quaternary ammonium groups.

Other, pharmaceutically acceptable salts of the compounds of Formulae V-VIII include acid addition salts such as those containing sulfate, chloride, hydrochloride, fumarate, maleate, phosphate, sulfamate, acetate, citrate, lactate, tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluene sulfonate, cyclohexylsulfamate and quinate. Pharmaceutically acceptable salts of the compounds of Formulae V-VIII can be obtained from acids such as hydrochloric acid, maleic acid, sulfuric acid, phosphoric acid, sulfamic acid, acetic acid, citric acid, lactic acid, tartaric acid, malonic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclohexylsulfamic acid, fumaric acid, and quinic acid.

Pharmaceutically acceptable salts of the compounds of Formulae V-VIII also include basic addition salts such as those containing benzathine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine, procaine, aluminum, calcium, lithium, magnesium, potassium, sodium, ammonium, alkylamine, and zinc, when acidic functional groups, such as carboxylic acid or phenol are present. For example, see Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Co., Easton, Pa., Vol. 2, p. 1457, 1995. Such salts of the compounds of Formulae V-VIII can be prepared using the appropriate corresponding bases.

Salts of the compounds of Formulae V-VIII can be prepared, for example, by dissolving the free-base form of a compound in a suitable solvent, such as an aqueous or aqueous-alcohol in solution containing the appropriate acid and then isolated by evaporating the solution. In another example, a salt is prepared by reacting the free base and acid in an organic solvent.

The salts of the compounds of Formulae V-VIII may be present as a complex. Examples of complexes include 8-chlorotheophylline complex (analogous to, e.g., dimenhydrinate: diphenhydramine 8-chlorotheophylline (1:1) complex; Dramamine) and various cyclodextrin inclusion complexes.

Solvents useful in the preparation of pharmaceutically acceptable salts of the compounds of Formulae V-VIII include organic solvents, such as for example, acetonitrile, acetone, alcohols (e.g., methanol, ethanol and isopropanol), tetrahydrofuran, methyl ethyl ketone (MEK), ethers (e.g., diethyl ether), benzene, toluene, xylenes, dimethylformamide (DMF), and N-methylpyrrolidinone (NMP), and the like. Preferably, the solvents are selected from acetonitrile and MEK.

The salts of compounds of Formulae V-VIII may be present as a complex. Examples of complexes include 8-chlorotheophylline complex (analogous to, e.g., dimenhydrinate: diphenhydramine 8-chlorotheophylline (1:1) complex; Dramamine) and various cyclodextrin inclusion complexes.

Formulations of salts of the compounds of Formulae I-VIII provided herein exhibit at least one, or preferably some or even more preferably, all the following characteristics: (1) they are stable at ambient temperatures for a minimum of one year; (2) they provide for ease of oral administration; (3) they facilitate patient compliance with dosing; (4) upon administration, they consistently facilitate high levels of absorption of the pharmaceutical active; (5) upon once or twice daily oral administration they allow release of the KATP channel opener over a sustained time frame such that the circulating concentration of the KATP channel opener or its metabolically active metabolites does not fall below a therapeutically effective concentration; (6) they achieve these results independent of the pH of the gastrointestinal tract of treated subjects, and (7) they delay release until gastric transit is complete or nearly complete.

Formulations designed for oral administration of the salts of the compounds of Formulae I-VIII can be provided, for example, as capsules, tablets, or as quick dissolve tablets or films. Capsule or tablet formulations include a number of distinguishing components. One is a component to improve absorption of the KATP channel opener. Another sustains release of the drug over more than 2 hours. A third delays substantial release of the drug until gastric transit is completed.

Oral administration formulations of the salts of the compounds of Formulae I-VIII can also be provided, for example, as oral suspensions, oral solutions, encapsulated oral suspensions, and encapsulated oral solutions. Formulations can be designed for immediate release or controlled release. Preferably, such oral formulations are not produced from a liquid form of the sodium salt of diazoxide.

Formulations of the salts of the compounds of Formulae I-VIII can also be prepared for transdermal, intranasal and intravenous (I.V.) administration, provided that when the anion is diazoxide and the cation is sodium, the formulation is not for intravenous use.

In another embodiment, formulations of the salts of the compounds of Formulae I-VIII are prepared for transdermal or intranasal, administration, provided that when the anion is diazoxide and the cation is sodium, the formulation is not produced using a liquid form of the salt of the compounds of Formulae I-VIII.

In another embodiment, formulations of the salts of the compounds of Formulae I-VIII are prepared for transdermal, intranasal and intravenous (I.V.) administration excluding the sodium salt of diazoxide.

Formulations of KATP channel openers prepared using salts of the compounds selected from Formulae I-VIII exhibit improved solubility and absorption compared to previous formulations of these drugs. These advantageous properties are achieved by any one or more of the following approaches: (1) reducing particle size of the formulation by comminution, spray drying, or other micronising techniques, (2) using an ion exchange resin in the formulation, (3) using inclusion complexes, for example using a cyclodextrin, (4) compaction of the salt of KATP channel opener with a solubilizing agent including low viscosity hypromellose, low viscosity methylcellulose or similarly functioning excipient and combinations thereof, (5) associating the salt of the KATP channel opener with a distinct salt prior to formulation, (6) using a solid dispersion of the salt of the KATP channel opener, (7) using a self emulsifying system, (8) adding one or more surfactants to the formulation, (9) using nanoparticles in the formulation, or (10) combinations of these approaches.

Release of KATP channel opener selected from salts of the compounds of Formulae I-VIII over a sustained period of time (e.g., 2-30 hours) can be achieved by the use of one or more approaches including, but not limited to: (1) the use of pH sensitive polymeric coatings, (2) the use of a hydrogel, (3) the use of a film coating that controls the rate of diffusion of the drug from a coated matrix, (4) the use of an erodable matrix that controls rate of drug release, (5) the use of polymer coated pellets, granules, or microparticles which can be further encapsulated or compressed into a tablet, (6) the use of an osmotic pump system, (7) the use of a compression coated tablet, or (8) combinations of these approaches.

Delay of release of KATP channel openers selected from the salts of the compounds of Formulae I-VIII from the formulation until gastric transit is complete can be achieved in the formulations provided herein by any of several mechanisms. For example, pH sensitive polymer or co-polymer can be used which when applied around the drug matrix functions as an effective barrier to release of active at pH 3.0 or lower and is unstable at pH 5.5 and above. This provides for control of release of the active compound in the stomach but rapidly allows release once the dosage form has passed into the small intestine. An alternative to a pH sensitive polymer or co-polymer is a polymer or co-polymer that is non-aqueous-soluble. The extent of resistance to release in the gastric environment can be controlled by coating with a blend of the non-aqueous-soluble and a aqueous soluble polymer. In this approach neither of the blended polymers or co-polyers are pH sensitive. One example of a pH sensitive co-polymer is the Eudragit® methacrylic co-polymers, including Eudragit® L 100, S 100 or L 100-55 solids, L 30 D-55 or FS 30D dispersions, or the L 12,5 or S 12,5 organic solutions.

Polymers that delay release can be applied to a tablet either by spray coating (as a thin film) or by compression coating. If a capsule is used, then the polymer(s) may be applied over the surface of the capsule or applied to microparticles of the drug, which may then be encapsulated such as in a capsule or gel. If the capsule is coated, then it will resist disintegration until after gastric transit. If microparticles are coated, then the capsule may disintegrate in the stomach but little to no drug will be released until after the free microparticles complete gastric transit. Finally, an osmotic pump system that uses e.g., a swellable hydrogel can be used to delay drug release in the stomach. The swellable hydrogel takes up moisture after administration. Swelling of the gel results in displacement of the drug from the system for absorption. The timing and rate of release of the drug depend on the gel used, and the rate at which moisture reaches the gel, which can be controlled by the size of the opening in the system through which fluid enters. See Drug Delivery Technologies online article Dong et al., “L-OROS® SOFTCAP™ for Controlled Release of Non-Aqueous Liquid Formulations.”

Accordingly, delay of release of formulations of KATP channel openers prepared as salts of the compounds of Formulae I-VIII until after gastric transit is complete can be achieved by any of several mechanisms, including, but not limited to: (a) a pH sensitive polymer or co-polymer applied as a compression coating on a tablet; (b) a pH sensitive polymer or co-polymer applied as a thin film on a tablet; (c) a pH sensitive polymer or co-polymer applied as a thin film to an encapsulation system; (d) a pH sensitive polymer or co-polymer applied to encapsulated microparticles, (e) a non-aqueous-soluble polymer or copolymer applied as a compression coating on a tablet; (f) a non-aqueous-soluble polymer or co-polymer applied as a thin film on a tablet; (g) a non-aqueous soluble polymer applied as a thin film to an encapsulation system; (h) a non-aqueous soluble polymer applied to microparticles; (i) incorporation of the formulation in an osmotic pump system, or (j) use of systems controlled by ion exchange resins, or (k) combinations of these approaches, wherein the pH sensitive polymer or co-polymer is resistant to degradation under acid conditions.

Formulations are provided that are designed for administration once daily (i.e., once per 24 hours). These formulations can contain between 25 and 500 mg of KATP channel openers selected from salts of the compounds of Formulae I-VIII. Formulations intended for administration twice daily (per 24 hours) may also be provided. These can contain between 25 and 250 mg of KATP channel openers.

The formulations provided herein exhibit improved safety of the administered drug product. This improvement in safety occurs by at least two mechanisms. First, delay of release of active drug until gastric transit is complete can reduce the incidence of a range of gastrointestinal adverse side effects including nausea, vomiting, dyspepsia, abdominal pain, diarrhea and ileus. Second, by sustaining release of the active drug over 2 or more hours up to as long as 24 hours, peak drug levels are reduced relative to the peak drug levels observed for the same administered dose using any oral formulation that does not have sustained or controlled release. This reduction in peak drug levels can contribute to reductions in adverse effects that are partially or completely determined by peak drug levels. These adverse effects include: fluid retention with the associated reduced rates of excretion of sodium, chloride and uric acid, edema, hyperglycemia and the associated potential for progression to ketoacidosis, cataracts and non-ketotic hyperosmolar coma, headaches, tachycardia and palpitations.

Also provided herein are controlled release formulations of KATP channel openers prepared from salts of compounds of Formulae I-VIII, which have one feature from each of A-D as shown in Table 1.

TABLE 1 Controlled Release Formulation Characteristics and Properties A. Unit Form: Tablet or Capsule B. Dosage/unit: 10-100 mg 100-200 mg 200-300 mg 300-500 mg 500-2000 mg C. Dosing Once daily (24 hours) Twice daily (24 hours) D. Release time: 2-4 hrs 4-8 hrs 8-24 hours

For example, a controlled release composition can be a tablet containing 25-100 mg of a salt of a compound of Formulae I-VIII, wherein such tablet administered once daily to achieve a controlled release time of 2-4 hours. All of these formulations can further include the feature of substantially delaying pharmaceutical active release until after gastric transit is complete.

In addition, any of the above formulations from Table 1 can include at least one feature that improves the solubility or absorption of the KATP channel opener.

Exemplary controlled release formulations provided herein include the active compound (I.e., a KATP channel opener selected from a salt of a compound of any of Formulae I-VIII) and a matrix which includes a gelling agent that swells upon contact with aqueous fluid. The active compound entrapped within the gel is slowly released into the body upon dissolution of the gel. The active compound can be evenly dispersed within the matrix or can be present as pockets of drug in the matrix. For example, the drug can be formulated into small granules which are dispersed within the matrix. In addition, the granules of drug also can include a matrix, thus, providing a primary and a secondary matrix as described in U.S. Pat. No. 4,880,830 to Rhodes.

The gelling agent preferably is a polymeric material, which can include, for example, any pharmaceutically acceptable water soluble or water insoluble slow releasing polymer such as xantham gum, gelatin, cellulose ethers, gum arabic, locust bean gum, guar gum, carboxyvinyl polymer, agar, acacia gum, tragacanth, veegum, sodium alginate or alginic acid, polyvinylpyrrolidone, polyvinyl alcohol, or film forming polymers such as methyl cellulose (MC), carboxymethyl cellulose (CMC), hydroxypropyl methylcellulose, hyroxypropyl methyl cellulose (HPMC), hydroxypropyl cellulose (HPC), hydroxyethyl cellulose (HEC), ethylcellulose (EC), acrylic resins or mixtures of the above (see e.g., U.S. Pat. No. 5,415,871).

The gelling agent of the matrix also can be a heterodisperse gum comprising a heteropolysaccharide component and a homopolysaccharide component which produces a fast-forming and rigid gel as described in U.S. Pat. No. 5,399,359. The matrix also can include a cross-linking agent such as a monovalent or multivalent metal cations to further add rigidity and decrease dissolution of the matrix, thus further slowing release of drug. The amount of crosslinking agent to add can be determined using methods routine to the ordinary skilled artisan.

The matrix of the controlled release composition also can include one or more pharmaceutically acceptable excipients recognized by those skilled in the art, i.e. formulation excipients. Such excipients include, for example, binders: polyvinylpyrrolidone, gelatin, starch paste, microcrystalline cellulose; diluents (or fillers): starch, sucrose, dextrose, lactose, fructose, xylitol, sorbitol, sodium chloride, dextrins, calcium phosphate, calcium sulphate; and lubricants: stearic acid, magnesium stearate, calcium stearate, Precirol® and flow aids for example talc or colloidal silicon dioxide.

The matrix of the controlled release composition can further include a hydrophobic material which slows the hydration of the gelling agent without disrupting the hydrophilic nature of the matrix, as described in U.S. Pat. No. 5,399,359. The hydrophobic polymer can include, for example, alkylcellulose such as ethylcellulose, other hydrophobic cellulosic materials, polymers or copolymers derived from acrylic or methacrylic acid esters, copolymers of acrylic and methacrylic acid esters, zein, waxes, shellac, hydrogenated vegetable oils, waxes and waxy substances such as carnauba wax, spermaceti wax, candellila wax, cocoa butter, cetosteryl alcohol, beeswax, ceresin, paraffin, myristyl alcohol, stearyl alcohol, cetylalcohol and stearic acid, and any other pharmaceutically acceptable hydrophobic material known to those skilled in the art.

The amount of hydrophobic material incorporated into the controlled release composition is that which is effective to slow the hydration of the gelling agent without disrupting the hydrophilic matrix formed upon exposure to an environmental fluid. In certain preferred embodiments, the hydrophobic material is included in the matrix in an amount from about 1 to about 20 percent by weight and replaces a corresponding amount of the formulation excipient. A solvent for the hydrophobic material may be an aqueous or organic solvent, or mixtures thereof.

Examples of commercially available alkylcelluloses are Aquacoat® (aqueous dispersion of ethylcellulose available from FMC) and Surelease® (aqueous dispersion of ethylcellulose available from Colorcon). Examples of commercially available acrylic polymers suitable for use as the hydrophobic material include Eudragit® RS and RL (copolymers of acrylic and methacrylic acid esters having a low content (e.g., 1:20 or 1:40) of quaternary ammonium compounds).

The controlled release composition also can be coated to retard access of liquids to the active compound and/or retard release of the active compound through the film-coating. The film-coating can provide characteristics of gastroresistance and enterosolubility by resisting rapid dissolution of the composition in the digestive tract. The film-coating generally represents about 5-15% by weight of the controlled release composition. Preferably, the core by weight represents about 90% of the composition with the remaining 10% provided by the coating. Such coating can be a film-coating as is well known in the art and include gels, waxes, fats, emulsifiers, combination of fats and emulsifiers, polymers, starch, and the like.

Polymers and co-polymers are useful as thin film coatings. Solution coatings and dispersion coatings can be used to coat the active compound, either alone or combined with a matrix. The coating is preferably applied to the drug or drug and matrix combination as a solid core of material as is well known in the art.

A solution for coating can include polymers in both organic solvent and aqueous solvent systems, and typically further including one or more compounds that act as a plasticizer. Polymers useful for coating compositions include, for example, methylcellulose (Methocel® A; Dow Chemical Co.), hydroxypropylmethylcellulose with a molecular weight between 1,000 and 4,000,000 (Methocel® E; Dow Chemical Co. or Pharmacoat®; Shin Etsu), hydroxypropyl cellulose with a molecular weight between 2,000 and 2,000,000, ethyl cellulose, cellulose acetate, cellulose triacetate, cellulose acetate butyrate, cellulose acetate phthalate, cellulose acetate trimellitate (Eastman Kodak), carboxymethylethyl cellulose (Duodcel®), hydroxypropyl methylcellulose phthalate, ethylcellulose, methylcellulose and, in general, cellulosic derivatives, polymethacrylic acid-methacrylic acid copolymer (Type A 1:1 Eudragit L100; Type B 1:2 Eudragit S100; and Type C 1:1 Eudragit L100-55, aqueous dispersion 30% solids, Eudragit L30D), poly(meth)acryl ester: poly(ethyl acrylate, methyl methacrylate 2:1), Eudragit NE30D aqueous dispersion 30% solids, polyaminomethacrylate Eudragit E100, poly(trimethylammonioethyl methacrylate chloride) ammoniomethacrylate copolymer, Eudragit RL30D and Eudragit RS30D, carboxyvinyl polymers, polyvinylalcohols, glucans scleroglucans, mannans, and xanthans.

Aqueous polymeric dispersions include Eudragit L30D and RS/RL30D, and NE30D, Aquacoat® brand ethyl cellulose, Surelease brand ethyl cellulose, EC brand N-10F ethyl cellulose, Aquateric brand cellulose acetate phthalate, Coateric brand Poly(vinyl acetate phthalate), and Aqacoat brand hydroxypropyl methylcellulose acetate succinate. Most of these dispersions are latex, pseudolatex powder or micronized powder mediums.

A plasticizing agent may be included in the coating to improve the elasticity and the stability of the polymer film and to prevent changes in the polymer permeability over prolonged storage. Such changes may affect the drug release rate. Suitable conventional plasticizing agents include, for example, diethyl phthalate, glycerol triacetate, acetylated monoglycerides, acetyltributylcitrate, acetyltriethyl citrate, castor oil, citric acid esters, dibutyl phthalate, dibutyl sebacate, diethyloxalate, diethyl malate, diethylfumarate, diethylphthalate, diethylsuccinate, diethylmalonate, diethyltartarate, dimethylphthalate, glycerin, glycerol, glyceryl triacetate, glyceryltributyrate, mineral oil and lanolin alcohols, petrolatum and lanolin alcohols, phthalic acid esters, polyethylene glycols, propylene glycol, rape oil, sesame oil, triacetin, tributyl citrate, triethyl citrate, and triethyl acetyl citrate, or a mixture of any two or more of the foregoing. Plasticizers which can be used for aqueous coatings include, for example, propylene glycol, polyethylene glycol (PEG 400), triacetin, polysorbate 80, triethyl citrate, and diethyl d-tartrate.

A coating solution comprising a mixture of hydroxypropylmethylcellulose and aqueous ethylcellulose (e.g. Aquacoat brand) as the polymer and dibutyl sebacate as plasticizer can be used for coating microparticles. (Aquacoat is an aqueous polymeric dispersion of ethylcellulose and contains sodium lauryl sulfate and cetyl alcohol). Preferably, the plasticizer represents about 1-2% of the composition.

In addition to the polymers, the coating layer can include an excipient to assist in formulation of the coating solution. Such excipients may include a lubricant or a wetting agent. Suitable lubricants as excipients for the film coating include, for example, talc, calcium stearate, colloidal silicon dioxide, glycerin, magnesium stearate, mineral oil, polyethylene glycol, and zinc stearate, aluminum stearate or a mixture of any two or more of the foregoing. Suitable wetting agents include, for example, sodium lauryl sulfate, acacia, benzalkonium chloride, cetomacrogol emulsifying wax, cetostearyl alcohol, cetyl alcohol, cholesterol, diethanolamine, docusate sodium, sodium stearate, emulsifying wax, glyceryl monostearate, hydroxypropyl cellulose, lanolin alcohols, lecithin, mineral oil, monoethanolamine, poloxamer, polyoxyethylene alkyl ethers, polyoxyethylene castor oil derivatives, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, propylene glycol alginate, sorbitan esters, stearyl alcohol and triethanolamine, or a mixture of any two or more of the foregoing.

The specified tablet or capsule formulations of Table 1 may include co-formulation with an obesity treating drug (in addition to a KATP channel opener selected from a salt of a compound of Formulae I-VIII). Obesity treating drugs that may be used include, but are not limited to, sibutramine hydrochloride (5-30 mg/unit), orlistat (50-360 mg/unit), phentermine hydrochloride or resin complex (15 to 40 mg/unit), zonisamide (100 to 600 mg/unit), topiramate (64 to 400 mg/unit), naltrexone hydrochloride (50 to 600 mg/unit), rimonabant (5 to 20 mg/unit), ADP356 (5 to 25 mg/unit), ATL962 (20 to 400 mg/unit), or AOD9604 (1 to 10 mg/unit). These formulations are preferably used once daily. For a twice daily dosing, the amount of KATP channel opener selected from a salt of a compound of Formulae I-VIII is one half the amount included in the once daily formulation and the co-formulated obesity treating drug is half of the amount specified. Alternative obesity treating drugs may include, but are not limited to: selective serotonin 2c receptor agonists, dopamine antagonists, cannabinoid-1 receptor antagonists, leptin analogues, leptin transport and/or leptin receptor promoters, neuropeptide Y and agouti-related peptide antagonists, proopiomelanocortin and cocaine and amphetamine regulated transcript promoters, melanocyte-stimulating hormone analogues, melanocortin-4 receptor agonists, and agents that affect insulin metabolism/activity, which can include protein-tyrosine phosphatase-1B inhibitors, peroxisome proliferator activated receptor antagonists, short-acting bromocriptine (ergoset), somatostatin agonists (octreotide), and adiponectin, gastrointestinal-neural pathway agents, including those that increase cholecystokinin activity, increase glucagon-like peptide-1 activity (e.g., exendin 4, liraglutide, dipeptidyl peptidase IV inhibitors), and increase protein YY3-36 activity and those that decrease ghrelin activity, as well as amylin analogues, agents that may increase resting metabolic rate (“selective” β-3 stimulators/agonist, uncoupling protein homologues, and thyroid receptor agonists), melanin concentrating hormone antagonists, phytostanol analogues, amylase inhibitors, growth hormone fragments, synthetic analogues of dehydroepiandrosterone sulfate, antagonists of adipocyte 11B hydroxysteroid dehydrogenase type 1 activity, corticotropin releasing hormone agonists, inhibitors of fatty acid synthesis, carboxypeptidase inhibitors, indanones/indanols, aminosterols, and other gastrointestinal lipase inhibitors.

The specified tablet or capsule formulations of Table 1 may include co-formulation with a diabetes treating drug (in addition to a KATP channel opener selected from a salt of a compound of Formulae I-VIII). Diabetes treating drugs that may be used include, but are not limited to, acarbose (50 to 300 mg/unit), miglitol (25 to 300 mg/unit), metformin hydrochloride (300 to 2000 mg/unit), repaglinide (1-16 mg/unit), nateglinide (200 to 400 mg/unit), rosiglitazone (5 to 50 mg/unit), metaglidasen (100 to 400 mg/unit) or any drug that improves insulin sensitivity, or improves glucose utilization and uptake. These formulations are preferably used once daily. For a twice daily dosing, the amount of the KATP channel opener selected from a salt of a compound of Formulae I-VIII is half the amount included in the once daily formulation and the co-formulated diabetes treating drug is half of the amount specified.

The specified tablet or capsule formulations of Table 1 may include co-formulation with a cholesterol lowering drug. Cholesterol lowering drugs that may be used include, but are not limited to, pravastatin, simvastatin, atorvastatin, fluvastatin, rosuvastatin, or lovastatin (all at 10 to 80 mg/unit), fibrates (50 to 300 mg/unit), niacin (500 to 2000 mg/unit), thyroid receptor activators (0.5 to 100 mg/unit), MTP inhibitors (20 to 1000 mg/unit), PPAR delta agonists and modulators (5 to 400 mg/unit), and squalene synthase inhibitor (10 to 1000 mg/unit). These formulations are preferably used once daily. Each of these classes of active may also be used to raise HDL cholesterol. For a twice daily dosing, the amount of KATP channel opener selected from a salt of a compound of Formulae I-VIII is preferably 25 to 200 mg/unit and the co-formulated cholesterol lowering drug is half of the amount specified.

The specified tablet or capsule formulations of Table 1 may include co-formulation with a depression treating drug. Depression treating drugs that may be used include, but are not limited to, citalopram hydrobromide (10 to 80 mg/unit), escitalopram hydrobromide (5 to 40 mg/unit), fluvoxamine maleate (25 to 300 mg/unit), paroxetine hydrochloride (12.5 to 75 mg/unit), fluoxetine hydrochloride (30 to 100 mg/unit), setraline hydrochloride (25 to 200 mg/unit), amitriptyline hydrochloride (10 to 200 mg/unit), desipramine hydrochloride (10 to 300 mg/unit), nortriptyline hydrochloride (10 to 150 mg/unit), duloxetine hydrochloride (20 to 210 mg/unit), venlafaxine hydrochloride (37.5 to 150 mg/unit), phenelzine sulfate (10 to 30 mg/unit), bupropion hydrochloride (200 to 400 mg/unit), or mirtazapine (7.5 to 90 mg/unit). These formulations are preferably used once daily. For a twice daily dosing, the amount of KATP channel opener selected from a salt of a compound of Formulae I-VIII is preferably half the amount included in the once daily formulation and the co-formulated depression treating drug is half of the amount specified.

The specified tablet or capsule formulations of Table 1 may include co-formulation with a hypertension treating drug. Hypertension treating drugs that may be used include, but are not limited, to enalapril maleate (2.5 to 40 mg/unit), captopril (2.5 to 150 mg/unit), lisinopril (10 to 40 mg/unit), benzaepril hydrochloride (10 to 80 mg/unit), quinapril hydrochloride (10 to 80 mg/unit), peridopril erbumine (4 to 8 mg/unit), ramipril (1.25 to 20 mg/unit), trandolapril (1 to 8 mg/unit), fosinopril sodium (10 to 80 mg/unit), moexipril hydrochloride (5 to 20 mg/unit), losartan potassium (25 to 200 mg/unit), irbesartan (75 to 600 mg/unit), valsartan (40 to 600 mg/unit), candesartan cilexetil (4 to 64 mg/unit), olmesartan medoxamil (5 to 80 mg/unit), telmisartan (20 to 160 mg/unit), eprosartan mesylate (75 to 600 mg/unit), atenolol (25 to 200 mg/unit), propranolol hydrochloride (10 to 180 mg/unit), metoprolol tartrate, succinate or fumarate (each at 25 to 400 mg/unit), nadolol (20 to 160 mg/unit), betaxolol hydrochloride (10 to 40 mg/unit), acebutolol hydrochloride (200 to 800 mg/unit), pindolol (5 to 20 mg/unit), bisoprolol fumarate (5 to 20 mg/unit), nifedipine (15 to 100 mg/unit), felodipine (2.5 to 20 mg/unit), amlodipine besylate (2.5 to 20 mg/unit), nicardipine (10 to 40 mg/unit), nisoldipine (10 to 80 mg/unit), terazosin hydrochloride (1 to 20 mg/unit), doxaxosin mesylate (4 to 16 mg/unit), prazosin hydrochloride (2.5 to 10 mg/unit), or alfuzosin hydrochloride (10 to 20 mg/unit). These formulations are preferably used once daily. For a twice daily dosing, the amount of KATP channel opener is preferably half the amount included in the once daily formulation and the co-formulated hypertension treating drug is half of the amount specified.

The specified tablet or capsule formulations of Table 1 may include co-formulation with a diuretic to treat edema. Diuretics that may be used include, but are not limited to amiloride hydrochloride (1 to 10 mg/unit), spironolactone (10 to 100 mg/unit), triamterene (25 to 200 mg/unit), bumetanide (0.5 to 4 mg/unit), furosemide (10 to 160 mg/unit), ethacrynic acid or ethacrynate sodium (each at 10 to 50 mg/unit), torsemide (5 to 100 mg/unit), chlorthalidone (10 to 200 mg/unit), indapamide (1 to 5 mg/unit), hydrochlorothiazide (10 to 100 mg/unit), chlorothiazide (50 to 500 mg/unit), bendroflumethiazide (5 to 25 mg/unit), hydroflumethiazide (10 to 50 mg/unit), mythyclothiazide (1 to 5 mg/unit), or polythiazide (1 to 10 mg/unit). These formulations are preferably used once daily. For a twice daily dosing, the amount of KATP channel opener selected from a salt of a compound of Formulae I-VIII is preferably half the amount included in the once daily formulation and the co-formulated diuretic is half of the amount specified.

The specified tablet or capsule formulations of Table 1 may include co-formulation with a drug to treat inflammation or pain. Drugs for treating inflammation or pain that may be used include, but are not limited to aspirin (100 to 1000 mg/unit), tramadol hydrochloride (25 to 150 mg/unit), gabapentin (100 to 800 mg/unit), acetaminophen (100 to 1000 mg/unit), carbamazepine (100 to 400 mg/unit), ibuprofen (100 to 1600 mg/unit), ketoprofen (12 to 200 mg/unit), fenprofen sodium (100 to 600 mg/unit), flurbiprofen sodium or flurbiprofen (both at 50 to 200 mg/unit), or combinations of any of these with a steroid or aspirin. These formulations are preferably used once daily. For a twice daily dosing, the amount of KATP channel opener selected from a salt of a compound of Formulae I-VIII is preferably half the amount included in the once daily formulation and the co-formulated diuretic is half of the amount specified.

The specified tablet or capsule formulations of Table 1 may include co-formulation with a drug to treat obesity associated co-morbidities include those specified above for treating diabetes, cholesterol, depression, hypertension and edema, or drugs to treat atherosclerosis, osteoarthritis, disc herniation, degeneration of knees and hips, breast, endometrial, cervical, colon, leukemia and prostate cancers, hyperlipidemia, asthma/reactive airway disease, gallstones, GERD, obstructive sleep apnea, obesity hypoventilation syndrome, recurrent ventral hernias, menstrual irregularity and infertility.

The specified tablet or capsule formulations of Table 1 may include co-formulation with an anti-psychotic drug. The combination used to treat the psychotic condition and to treat or prevent weight gain, dyslipidemia or impaired glucose tolerance in the treated subject. Drugs for treating various psychotic conditions that may be used include, but are not limited to, lithium or a salt thereof (250 to 2500 mg/unit), carbamazepine or a salt thereof (50 to 1200 mg/unit), valproate, valproic acid, or divalproex (125 to 2500 mg/unit), lamotrigine (12.5 to 200 mg/unit), olanzapine (5 to 20 mg/unit), clozapine (12.5 to 450 mg/unit), or risperidone (0.25 to 4 mg/unit). These coformulations are preferably intended for once per day administration. For a twice daily dosing, the amount of KATP channel opener selected from a salt of a compound of Formulae I-VIII is preferably half the amount included in the once daily formulation and the co-formulated anti-psychotic is half of the amount specified.

The specified tablet or capsule formulations of Table 1 may include co-formulation with a drug to treat or prevent ischemic or reperfusion injury. Drugs for treating or preventing ischemic or reperfusion injury that may be used include, but are not limited to: low molecular weight heparins (e.g., dalteparin, enoxaparin, nadroparin, tinzaparin or danaparoid), ancrod, pentoxifylline, nimodipine, flunarizine, ebselen, tirilazad, clomethiazole, an AMPA agonist (e.g., GYKI 52466, NBQX, YM90K, zonampanel, or MPQX), SYM 2081, selfotel, Cerestat, CP-101,606, dextrophan, dextromethorphan, MK-801, NPS1502, remacemide, ACEA 1021, GV150526, eliprodil ifenprodil, lubeluzole, naloxone, nalmefene citicoline, acetyl-1-carnitine, nifedipine, resveratrol, a nitrone derivative, clopidogrel, dabigatram, prasugrel, troxoprodil, AGY-94806, or KAI-9803.

Provided are formulations administered once or twice daily to an obese or overweight subject continuously result in a circulating concentration of KATP channel opener selected from a salt of a compound of Formulae I-VIII sufficient to induce weight loss. Weight loss occurs by the preferential loss of body fat. Additional weight loss can occur when the formulation is administered in combination with a reduced calorie diet.

Provided are formulations of KATP channel openers selected from a salt of a compound of Formulae I-VIII administered as a single dose to an obese, overweight or obesity-prone subject that result in the inhibition of fasting or glucose stimulated insulin secretion for about 24 hours or for about 18 hours.

Provided are formulations of KATP channel openers selected from a salt of a compound of Formulae I-VIII administered as a single dose to an obese, overweight or obesity-prone subject that result in the elevation of energy expenditure for about 24 hours or for about 18 hours.

Provided are formulations of KATP channel openers selected from a salt of a compound of Formulae I-VIII administered as a single dose to an obese, overweight or obesity-prone subject that result in the elevation of beta oxidation of fat for about 24 hours or for about 18 hours.

Provided are formulations of KATP channel openers selected from a salt of a compound of Formulae I-VIII administered as a single dose to an obese, overweight or obesity-prone hyperphagic subject that result in the inhibition of hyperphagia for about 24 hours or for about 18 hours.

Provided are formulations suitable for continuous administration once or twice daily (per 24 hours) to a subject, resulting in a circulating concentration of KATP channel openers selected from a salt of a compound of Formulae I-VIII sufficient to induce either beta-cell rest or improved insulin sensitivity or both. Such beta-cell rest and improvements in insulin sensitivity can contribute to effective treatment of type I diabetes, type II diabetes and prediabetes. Such beta-cell rest and improvements in insulin sensitivity can contribute to effective restoration of normal glucose tolerance in type II diabetic and prediabetic subjects.

The various pharmaceutical KATP channel opener formulations selected from a salt of a compound of Formulae I-VIII have a variety of applications, including, but not limited to: (1) treatment of obesity; (2) prevention of weight gain in subjects who are predisposed to obesity; (3) treatment of hyperinsulinemia or hyperinsulinism; (4) treatment of hypoglycemia; (5) treatment of hyperlipidemia, (6) treatment of type II diabetes, (7) preservation of pancreatic function in type I diabetics; (8) treatment of metabolic syndrome (or syndrome X); (9) prevention of the transition from prediabetes to diabetes, (10) correction of the defects in insulin secretion and insulin sensitivity contributing to prediabetes and type II diabetes, (11) treatment of polycystic ovary syndrome, (12) prevention of ischemic or reperfusion injury, (13) treat weight gain, dyslipidemia, or impairment of glucose tolerance in subjects treated with antipsychotics drugs, (14) prevent weight gain, dyslipidemia, or impairment of glucose tolerance in subjects treated with antipsychotics drugs and (15) treatment of any disease where hyperlipidemia, hyperinsulinemia, hyperinsulinism, hyperlipidemia, hyperphagia or obesity are contributing factors to the severity or progression of the disease, including but not limited to, Prader Willi Syndrome, Froelich's syndrome, Cohen syndrome, Summit Syndrome, Alstrom Syndrome, Borjesen Syndrome, Bardet-Biedl Syndrome, or hyperlipoproteinemia type I, II, III, and IV.

In one embodiment, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an overweight or obese subject as an oral dosage once per 24 hours to induce weight loss. In further embodiments, the subject (a) is not a type I diabetic, (b) is not a type II diabetic, (c) is not experiencing chronic, recurrent or drug-induced hypoglycemia, (d) does not have metabolic syndrome, or (e) is not experiencing malignant hypertension.

In one embodiment, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an overweight or obese subject as an oral dosage twice per 24 hours to induce weight loss. This treatment can be the sole treatment to induce weight loss. In further embodiments, the overweight or obese subject (a) does not have an insulin secreting tumor, (b) is not suffering from Poly Cystic Ovary Syndrome, (c) is not a type I diabetic, (d) is not a type II diabetic, (e) does not have metabolic syndrome, (f) is not experiencing chronic recurrent or drug-induced hypoglycemia, (g) has not been treated for schizophrenia with haloperidol, or (h) is not experiencing malignant hypertension. In further embodiments, the overweight or obese adolescent (a) has not been diagnosed as being type I or type II diabetic, (b) is not experiencing chronic, recurrent or drug-induced hypoglycemia, or (c) has not been diagnosed as having metabolic syndrome.

In another embodiment, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an overweight or obese subject as an oral dosage form three times per 24 hours to induce weight loss. This treatment can be the sole treatment to induce weight loss. In further embodiments, the overweight or obese subject (a) does not have an insulin-secreting tumor, (b) is not suffering from Poly Cystic Ovary Syndrome, (c) is not a type I diabetic, (d) is not a type II diabetic, (e) does not have metabolic syndrome, or (f) is not experiencing chronic, recurrent or drug-induced hypoglycemia.

In another embodiment, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an overweight or obese adolescent as an oral dosage form three times per 24 hours to induce weight loss. This treatment can be the sole treatment to induce weight loss. In further embodiments, the overweight or obese adolescent is (a) is not a type I or type II diabetic, (b) is not experiencing chronic, recurrent or drug-induced hypoglycemia or (c) does not have metabolic syndrome.

In another embodiment, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered as an oral dosage form three times per 24 hours to induce weight loss to an overweight or obese adult who (a) is not simultaneously receiving glucagon injections, triiodothyroxin or furosemide, (b) is not being treated for schizophrenia with haloperidol, or (c) is not experiencing malignant hypertension.

In another embodiment, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an overweight or obese subject as an oral dosage form four times per 24 hours to induce weight loss.

In another embodiment, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an overweight or obese subject as an oral dosage form administered from one, two, three or four times per 24 hours to induce weight loss at a daily dose of 50 to 700 mg. In a further embodiment, the overweight or obese subject (a) is not type I diabetic, (b) is not type II diabetic, (c) is not suffering chronic, recurrent or drug-induced hypoglycemia, or (d) does not have metabolic syndrome.

In another embodiment, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an overweight or obese subject as an oral dosage form administered from one, two, three or four times per 24 hours to induce weight loss at a daily dose of 130 to 400 mg. In a further embodiment, the overweight or obese subject (a) is not type I diabetic, (b) is not type II diabetic, (c) is not suffering chronic, recurrent or drug-induced hypoglycemia, or (d) does not have metabolic syndrome.

In other embodiments, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an overweight or obesity prone subject as an oral dosage form one, two, three or four times per 24 hours to maintain a weight loss, as it is preferable to maintain weight in an obese subject once some weight loss has occurred when the alternative is to regain weight. In a further embodiment, the administered daily dose of the KATP channel opener is 50 to 275 mg.

In other embodiments, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered as an oral dosage form to an overweight, obese, or obesity prone subject to (a) elevate energy expenditure, (b) elevate beta oxidation of fat, or (c) reduce circulating triglyceride concentrations.

In other embodiments, an oral dosage of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an subject in need thereof to induce the loss of 25%, 50%, or 75% of initial body fat.

In another embodiment, an oral dosage of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an subject in need thereof to induce (a) the preferential loss of body fat or (b) the preferential loss of visceral body fat.

In additional embodiments, an oral dosage of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered one, two or three times per 24 hours at daily doses of 50 to 700 mg to an subject to (a) induce the loss of 25%, 50% or 75% of initial body fat, (b) induce the preferential loss of body fat, or (c) induce the preferential loss of visceral fat.

In another embodiment, an oral dosage of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an subject to induce the preferential loss of body fat and to induce reduction in circulating triglycerides.

In another embodiment, an oral dosage of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is co-administered with sibutramine, orlistat, rimonabant, an appetite suppressant, an anti-depressant, an anti-epileptic, a diuretic, a drug that induces weight loss by a mechanism that is distinct from a KATP channel opener, or a drug that lowers blood pressure, to induce weight loss and/or treat obesity associated co-morbidities in an overweight, obese, or obesity prone subject. In further embodiments, the overweight, obese, or obesity prone subject (a) is a type I diabetic, (b) is not a type II diabetic, (c) is not suffering from chronic, recurrent or drug-induced hypoglycemia, or (d) does not have metabolic syndrome.

In another embodiment an oral dosage of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is co-administered with an anti-depressant, a drug that lowers blood pressure, a drug that lowers cholesterol, a drug that raises HDL, an anti-inflammatory that is not a Cox-2 inhibitor, a drug that lowers circulating triglycerides, to an overweight, obese, or obesity prone subject to induce weight loss and/or treat obesity associated co-morbidities. In further embodiments, the overweight, obese, or obesity prone subject (a) is not a type I diabetic, (b) is not a type II diabetic, (c) is not suffering from chronic, recurrent or drug-induced hypoglycemia, or (d) does not have metabolic syndrome.

In another embodiment, an oral dosage of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is co-administered with a drug that lowers blood pressure, a drug that lowers cholesterol, a drug that raises HDL, an anti-inflammatory that is not a Cox-2 inhibitor, a drug that lowers circulating triglycerides, to maintain weight and/or treat obesity associated co-morbidities in an overweight, obese, or obesity prone subject, as it is preferable to maintain weight in an obese subject once some weight loss has occurred when the alternative is to regain weight. In further embodiments, the overweight, obese, or obesity prone subject (a) is not a type I diabetic, (b) is not a type II diabetic, (c) is not suffering from chronic, recurrent or drug-induced hypoglycemia, or (d) does not have metabolic syndrome.

In additional embodiments, an oral dosage form of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is used to administer a therapeutically effective dose of a KATP channel opener to an obese, overweight or obesity prone subject in need thereof to treat obesity, to (a) provide beta cell rest, (b) treat type I or type II diabetes, or (c) prevent the occurrence of diabetes.

In additional embodiments, an oral dosage form of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is co-administered with Phentermine or a derivative thereof to an obese adult or adolescent to induce weight loss and/or treat obesity and obesity-associated co-morbidities. In further embodiments, a solid oral dosage form or tablet formulation of a KATP channel opener is co-administered with Phentermine or a derivative thereof to an obese adult or adolescent to treat metabolic syndrome in a patient in need thereof.

In further embodiments, a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII at doses of 50 to 700 mg/day is co-administered with Phentermine or a derivative thereof at daily doses of 15 to 37.5 mg to an overweight or obese subject to induce weight loss, to treat metabolic syndrome, or to induce weight loss and treat obesity-associated co-morbidities.

In another embodiment, a quick dissolving formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is used to provide a therapeutically effective dose to a patient in need thereof.

In further embodiments, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered once per 24 hours at doses of 50 mg to 700 mg to an overweight or obese subject.

In further embodiments, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is formulated as a tablet or capsule for oral administration. The tablet or capsule may be co-formulated with metformin. In another embodiment, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is formulated as an oral suspension or solution, and the oral suspension or solution may be further encapsulated in another embodiment.

In another embodiment, a pharmaceutical salt of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is formulated as a tablet or capsule for oral administration, or as an oral suspension or as an oral solution, or as an oral suspension or solution that is encapsulated.

In another embodiment a KATP channel opener selected from a salt of a compound of Formulae I-VIII is co-formulated with hydro-chlorothiazide, chlorothiazide, cyclothiazide, benzthiazide, metyclothiazide, bendro-flumethiazide, hydroflumethiazide, trichlormethiazide, or polythiazide in a pharmaceutical formulation suitable for oral administration.

Upon administration of formulations which include a salt of a compound of Formulae I-VIII provided herein to humans or animals, some or all of the following effects are observed: (1) the production of lipoprotein lipase by adipocytes is reduced; (2) enhanced lipolysis by adipocytes; (3) expression of fatty acid synthase by adipocytes is reduced; (4) glyceraldehydes phosphate dehydrogenase activity of adipocytes is reduced; (5) little or no new triglycerides are synthesized and stored by adipocytes; (6) enhanced expression of β3 Adrenergic Receptor (β3AR) an improvement in the adrenergic function in adipocytes; (7) reduced glucose stimulated secretion of insulin by pancreatic B-cells; (8) decreased insulinemia; (9) enhanced blood glucose levels; (10) increased expression of Uncoupling Protein 1 in adipocytes; (11) enhanced thermogenesis in white and brown adipose tissue; (12) reduction of plasma triglyceride concentration; (13) decrease in circulating leptin concentrations; (14) up-regulation of insulin receptors; (15) enhanced glucose uptake; (16) reduced adipocyte hyperplasia; (17) reduced adipocyte hypertrophy; (18) reduced rates of conversion of preadipocytes to adipocytes; (19) reduced rates of hyperphagia; (20) increased protection of CNS, cardiac and other tissues from ischemic or reperfusion injury; (21) improved insulin sensitivity; (22) elevated CSF insulin concentrations; (23) elevated circulating adiponectin concentrations; (25) reduced circulating triglyceride concentrations; (26) enhancement of beta-cell rest.

Threshold concentrations of the current invention include those circulating concentrations of KATP channel openers resulting from the administration of salts of compounds of Formulae I-VIII as an i.v. formulation, an immediate release oral formulation, a controlled release formulation, a transdermal formulation, or an intranasal formulation to an overweight or obese subject which results in (1) measurable suppression of fasting insulin levels, (2) suppression of fasting insulin levels by at least 20% from the baseline measurement in the same subject prior to treatment with a K_(ATP) channel opener selected from a salt of a compound of Formulae I-VIII, (3) suppression of fasting insulin levels by at least 30% from the baseline measurement in the same subject prior to treatment with a K_(ATP) channel opener selected from a salt of a compound of Formulae I-VIII, (4) suppression of fasting insulin levels by at least 40% from the baseline measurement in the same subject prior to treatment with a K_(ATP) channel opener selected from a salt of a compound of Formulae I-VIII, (5) suppression of fasting insulin levels by at least 50% from the baseline measurement in the same subject prior to treatment with a K_(ATP) channel opener selected from a salt of a compound of Formulae I-VIII, (6) suppression of fasting insulin levels by at least 60% from the baseline measurement in the same subject prior to treatment with a K_(ATP) channel opener selected from a salt of a compound of Formulae I-VIII, (7) suppression of fasting insulin levels by at least 70% from the baseline measurement in the same subject prior to treatment with a K_(ATP) channel opener selected from a salt of a compound of Formulae I-VIII, (8) suppression of fasting insulin levels by at least 80% from the baseline measurement in the same subject prior to treatment with a K_(ATP) channel opener selected from a salt of a compound of Formulae I-VIII, (9) loss of weight, (10) elevation of resting energy expenditure, or (11) elevation of the oxidation of fat or fatty acids. Threshold effects of the current invention include those circulating concentrations of K_(ATP) channel openers selected from salts of compounds of Formulae I-VIII resulting from the administration of an i.v. formulation of the drug, or an immediate release oral formulation of the drug, or a controlled release formulation of the drug, or a sustained release formulation, or a transdermal formulation, or an intranasal formulation of the drug to an obesity prone subject which result in (1) the loss of weight, and (2) the maintenance of weight. Threshold effects of the current invention include those circulating concentrations of K_(ATP) channel openers selected from salts of compounds of Formulae I-VIII resulting from the administration of an i.v. formulation of the drug, or an immediate release oral formulation of the drug, or a controlled release formulation of the drug, or a sustained release formulation, or a transdermal formulation, or an intranasal formulation of the drug to a prediabetic subject which result in prevention of the transition to diabetes. Threshold effects of the current invention include those circulating concentrations of K_(ATP) channel openers resulting from the administration of salts of compounds of Formulae I-VIII as an i.v. formulation, or an immediate release oral formulation, or a controlled release formulation, or a sustained release formulation, or a transdermal formulation, or an intranasal formulation to a subject with type 1 diabetes which result in beta cell rest.

The mode of action by which weight is maintained or lost resulting from the prolonged administration of K_(ATP) channel openers selected from salts of compounds of Formulae I-VIII to overweight, obese or obesity prone subjects as provided herein includes, but is not limited to, one or more of (1) enhanced energy expenditure, (2) enhanced oxidation of fat and fatty acids, (3) enhancement of lipolysis in adipose tissue, (4) enhanced glucose uptake by tissues and enhanced insulin sensitivity, and (5) improved beta adrenergic response. The mode of action by which weight is maintained or lost resulting from the prolonged administration of K_(ATP) channel openers selected from salts of compounds of Formulae I-VIII to obese or obesity prone subjects as provided herein may also include the suppression of appetite.

Prolonged administration of pharmaceutical formulations of KATP channel openers selected from salts of compounds of Formulae I-VIII to overweight or obese humans or animals results in substantial and sustained weight loss including some or all of the following effects: (1) preferential loss of body fat; (2) loss of greater than 25% of initial body fat mass; (3) loss of greater than 50% of initial body fat mass; (4) loss of greater than 75% of initial body fat mass; (5) significant increase in resting energy expenditure; (6) increase in the oxidation of fat and fatty acids; (7) reduction in blood pressure; (8) production of lipoprotein lipase by adipocytes is reduced; (9) enhanced lipolysis by adipocytes; (10) expression of fatty acid synthase by adipocytes is reduced; (11) glyceraldehydes phosphate dehydrogenase activity of adipocytes is reduced; (12) little or no new triglycerides are synthesized and stored by adipocytes; (13) enhanced expression of β3 Adrenergic Receptor (β3AR) and an improvement in the adrenergic function in adipocytes; (14) reduced glucose stimulated secretion of insulin by pancreatic B-cells; (15) decreased insulinemia; (16) enhanced blood glucose levels; (17) increased expression of Uncoupling Protein 1 in adipocytes; (18) enhanced thermogenesis in white and brown adipose tissue; (19) reduction of plasma triglyceride concentration; (20) decrease in circulating leptin concentrations; (21) up-regulation of insulin receptors; (22) enhanced glucose uptake; (23) reduced adipocyte hyperplasia; (24) reduced adipocyte hypertrophy; (25) reduced rates of conversion of preadipocytes to adipocytes; (26) reduced rates of hyperphagia; (27) the sequential loss first of the metabolically most active adipose tissue (visceral), followed by the loss of less metabolically active adipose tissue; (28) elevation of circulating adiponectin concentrations; (29) elevation of cerebrospinal fluid insulin levels; (30) enhanced islet insulin mRNA and insulin content; or (31) enhanced metabolic efficiency of insulin.

Prolonged administration of formulations of KATP channel openers selected from salts of compounds of Formulae I-VIII to obesity prone humans or animals, including subjects who have undergone various types of bariatric surgery, results in sustained maintenance of weight including some or all of the following effects: (1) increased resting energy expenditure; (2) increase in the oxidation of fat and fatty acids; (3) reduction in blood pressure; (4) production of lipoprotein lipase by adipocytes is reduced; (5) enhanced lipolysis by adipocytes; (6) expression of fatty acid synthase by adipocytes is reduced; (7) glyceraldehyde phosphate dehydrogenase activity of adipocytes is reduced; (8) little or no new triglycerides are synthesized and stored by adipocytes; (9) enhanced expression of β3 Adrenergic Receptor (β3AR) and improvement in the adrenergic function in adipocytes; (10) reduced glucose stimulated secretion of insulin by pancreatic B-cells; (11) decreased insulinemia; (12) enhanced blood glucose levels; (13) increased expression of Uncoupling Protein 1 in adipocytes; (14) enhanced thermogenesis in white and brown adipose tissue; (15) reduction of plasma triglyceride concentration; (16) decreased circulating leptin concentration; (17) up-regulation of insulin receptors; (18) enhanced glucose uptake; (19) reduced adipocyte hyperplasia; (20) reduced adipocyte hypertrophy; (21) reduced rates of conversion of preadipocytes to adipocytes; (22) reduced rates of hyperphagia; (23) elevated circulating adiponectin concentration; (24) elevated cerebrospinal fluid insulin levels; (25) enhanced islet insulin mRNA and insulin content; or (26) enhanced metabolic efficiency of insulin.

Immediate or prolonged administration of formulations of KATP channel openers selected from salts of compounds of Formulae I-VIII to prediabetic or type I diabetic humans or animals results in the prevention of beta cell failure, improved glycemic control, and prevention of the transition from prediabetes to diabetes including some or all of the following effects: (1) increase in resting energy expenditure; (2) increase in the oxidation of fat and fatty acids; (3) reduction in blood pressure; (4) production of lipoprotein lipase by adipocytes is reduced; (5) enhanced lipolysis by adipocytes; (6) expression of fatty acid synthase by adipocytes is reduced; (7) glyceraldehyde phosphate dehydrogenase activity of adipocytes is reduced; (8) little or no new triglycerides are synthesized and stored by adipocytes; (9) enhanced expression of β3 Adrenergic Receptor (β3AR) and an improvement in the adrenergic function in adipocytes; (10) reduced glucose stimulated secretion of insulin by pancreatic B-cells; (11) decreased insulinemia; (12) enhanced blood glucose levels; (13) increased expression of Uncoupling Protein 1 in adipocytes; (14) enhanced thermogenesis in white and brown adipose tissue; (15) reduction of plasma triglyceride concentration; (16) decreased circulating leptin concentrations; (17) up-regulation of insulin receptors; (18) enhanced glucose uptake; (19) reduced adipocyte hyperplasia; (20) reduced adipocyte hypertrophy; (21) reduced rates of conversion of preadipocytes to adipocytes; (22) reduced rates of hyperphagia; (23) elevated circulating adiponectin concentrations; (24) elevated cerebrospinal fluid insulin levels; (25) enhanced islet insulin mRNA and insulin content; or (26) enhanced metabolic efficiency of insulin.

Immediate or prolonged administration of formulations of KATP channel openers selected from salts of compounds of Formulae I-VIII to humans or animals that are at risk for myocardial infarct, or stroke, or undergoing surgical procedure that restores blood flow to heart or brain results in improved therapeutic outcomes post-surgically, or following the occurrence of myocardial infarct or stroke by improving the survival of tissue after blood flow is restored, reduced stunning of tissue, and altering the nature of the inflammatory responses.

Pharmaceutical formulations as provided herein are designed to be used in the treatment of obesity, hyperlipidemia, hypertension, weight maintenance, type I diabetes, prediabetes, type II diabetes, metabolic syndrome or any condition where weight loss, reduction in circulating triglycerides or beta cell rest contributes to therapeutic outcomes provide for a range of critical changes in pharmacodynamic and pharmacokinetic responses to administered doses of KATP channel openers selected from salts of compounds of Formulae I-VIII which changes include one or more of the following: (1) extending the pharmacodynamic effect of an administered dose to 24 hours or longer as measured by the suppression of insulin secretion; (2) providing for substantial uptake of the active pharmaceutical ingredient in the small intestine; (3) providing for substantial uptake of the active pharmaceutical ingredient in the large intestine; (4) result in lowered Cmax versus current oral suspension or capsule products for the same administered dose of active pharmaceutical ingredient; (5) provide for circulating concentrations of unbound active pharmaceutical ingredient above threshold concentrations for 24 or more hours from a single administered dose; and (6) provide for more consistent drug absorption by treated subjects as compared to existing capsule formulations.

Pharmaceutical co-formulations of the current invention designed to treat a range of conditions in humans and animals include the combination of KATP channel openers selected from salts of compounds of Formulae I-VIII with: (1) a diuretic, (2) a drug that lowers blood pressure, (3) a drug that suppresses appetite, (4) a cannabinoid receptor antagonist, (5) a drug that suppresses that action of gastric lipases, (6) any drug that is used to induce weight loss, (7) a drug that lowers cholesterol, (8) a drug that lowers LDL bound cholesterol, (9) a drug that improves insulin sensitivity, (10) a drug that improves glucose utilization or uptake, (11) a drug that reduces incidence of atherosclerotic plaque, (12) a drug that reduces inflammation, (13) a drug that is antidepressant, (14) a drug that is an anti-epileptic, or (15) a drug that is an anti-psychotic.

Treatment of humans or animals using pharmaceutical formulations (which include KATP channel openers selected from salts of compounds of Formulae I-VIII) result in reduced incidence of adverse side effects including but not limited to edema, fluid retention, reduced rates of excretion of sodium, chloride, and uric acid, hyperglycemia, ketoacidosis, nausea, vomiting, dyspepsia, ileus and headaches. These reductions in frequency of adverse side effects are achieved by: (1) initiating dosing of subjects at subtherapeutic doses and in a step wise manner increasing the dose daily until the therapeutic dose is achieved where the number of days over which the step up in dose is effected is 2 to 10, (2) use of the lowest effective dose to achieve the desired therapeutic effect, (3) use of a pharmaceutical formulation that delays release of active until gastric transit is complete, (4) use of a pharmaceutical formulation that results in lower circulating peak drug levels as compared to an immediate release oral suspension or capsule formulation for the same administered dose, and (5) optimizing the timing of administration of dose within the day and relative to meals.

Treatment of patients suffering from Prader Willi Syndrome, Froelich's Syndrome, Cohen Syndrome, Summit Syndrome, Alstrom Syndrome, Borjesen Syndrome, Bardet-Biedl Syndrome, and hyperlipoproteinemia type I, II, III, and IV with the current invention using pharmaceutical formulations of KATP channel openers selected from salts of compounds of Formulae I-VIII result in some or all of the following therapeutic outcomes: (1) weight loss, (2) reduced rates of weight gain, (3) inhibition of hyperphagia, (4) reduced incidence of impaired glucose tolerance, prediabetes or diabetes, (5) reduced incidence of congestive heart failure, (6) reduced hypertension, and (7) reduced rates of all cause mortality.

Treatment of prediabetic subjects using invention pharmaceutical formulations of KATP channel openers selected from salts of compounds of Formulae I-VIII result in some or all of the following therapeutic outcomes: (1) weight loss, (2) restoration of normal glucose tolerance, (3) delayed rates of progression to diabetes, (4) reduced hypertension, and (5) reduced rates of all cause mortality.

Treatment of diabetic subjects using invention pharmaceutical formulations of KATP channel openers selected from salts of compounds of Formulae I-VIII result in some or all of the following therapeutic outcomes: (1) weight loss, (2) restoration of normal glucose tolerance, (3) delayed rates of progression of diabetes, (4) improvements in glucose tolerance, (5) reduced hypertension, and (6) reduced rates of all cause mortality.

Co-administration of drugs with formulations of KATP channel openers selected from salts of compounds of Formulae I-VIII in the treatment of diseases of overweight, obese or obesity prone human and animal subjects involves the co-administration of a pharmaceutically acceptable formulation of KATP channel openers with an acceptable formulation of: (1) sibutramine, (2) orlistat, (3) rimonabant, (4) a drug that is an appetite suppressant, (5) any drug used to induce weight loss in an obese or overweight subject, (6) a non-thiazide diuretic, (7) a drug that lowers cholesterol, (8) a drug that raises HDL cholesterol, (9) a drug that lowers LDL cholesterol, (10) a drug that lowers blood pressure, (11) a drug that is an anti-depressant, (12) a drug that improves insulin sensitivity, (13) a drug that improves glucose utilization and uptake (14) a drug that is an anti-epileptic, (15) a drug that is an anti-inflammatory, or (16) a drug that lowers circulating triglycerides.

Co-administration of drugs with formulations of KATP channel openers selected from salts of compounds of Formulae I-VIII in the treatment or prevention of weight gain, dyslipidemia, impaired glucose tolerance or diabetes in subjects treated with antipsychotics drugs involve the co-administration of a pharmaceutically acceptable formulation of KATP channel openers with an acceptable formulation of: lithium, carbamazepine, valproic acid and divalproex, and lamotrigine; antidepressants generally classified as monoamine oxidase inhibitors including isocarboxazid, phenelzine sulfate and tranylcypromine sulfate; tricyclic antidepressants including doxepin, clomipramine, amitriptyline, maprotiline, desipromine, nortryptyline, desipramine, doxepin, trimipramine, imipramine and protryptyline; tetracyclic antidepressants including mianserin, mirtazapine, maprotiline, oxaprotiline, delequamine, levoprotiline, triflucarbine, setiptiline, azipramine, aptazapine maleate and pirlindole; and major tranquilizers and atypical antipsychotics including perphenazine, thioridazine, risperidone, clozapine, olanzapine and chlorpromazine.

In one embodiment, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an overweight or obese subject as an oral, transdermal or intranasal formulation to reach and maintain the threshold concentration required to measurably reduce fasting insulin levels for a prolonged period. Preferably the KATP channel opener formulation reduces fasting insulin levels by at least 20%, more preferably by at least 30%, more preferably by at least by 40%, more preferably by at least 50%, more preferably by at least by 60%, more preferably by at least by 70%, and more preferably by at least 80%. Fasting insulin levels are commonly measured using the glucose tolerance test (OGTT). After an overnight fast, a patient ingests a known amount of glucose. Initial glucose levels are determined by measuring pre-test glucose levels in blood and urine. Blood insulin levels are measured by a blood is draw every hour after the glucose is consumed for up to three hours. In a fasting glucose assay, subjects with plasma glucose values greater than 200 mg/dl at 2 hours post-glucose load indicate an impaired glucose tolerance.

In another embodiment, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an overweight or obese subject as an oral, transdermal or intranasal formulation to reach and maintain the threshold concentration required to induce weight loss for a prolonged period.

In another embodiment, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an overweight or obese subject as an oral, transdermal or intranasal formulation to reach and maintain the threshold concentration required to elevate resting energy expenditure for a prolonged period.

In another embodiment, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an overweight or obese subject as an oral, transdermal or intranasal formulation to reach and maintain the threshold concentration required to elevate fat and fatty acid oxidation for a prolonged period.

In another embodiment, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an obesity prone subject as an oral, transdermal or intranasal formulation to reach and maintain the threshold concentration required to induce weight loss for a prolonged period.

In another embodiment, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an obesity prone subject as an oral, transdermal or intranasal formulation to reach and maintain the threshold concentration required to maintain weight for a prolonged period.

In another embodiment, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an overweight or obese subject as an oral, transdermal or intranasal formulation to reach and maintain a drug concentration above the threshold concentration required to induce weight loss for a prolonged period.

In another embodiment, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an overweight or obese subject as an oral, transdermal or intranasal formulation for a prolonged period of time to reduce body fat by more than 25%, more preferably by at least 50%, and more preferably by at least 75%.

In another embodiment, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an overweight or obese subject as an oral, transdermal or intranasal formulation for a prolonged period of time to preferentially reduce visceral fat deposits.

In another embodiment, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an overweight or obese subject as an oral, transdermal or intranasal formulation for a prolonged period of time to reduce visceral fat depots and other fat deposits.

In another embodiment, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to a normoinsulinemic overweight or obese subject as an oral, transdermal or intranasal formulation to reach and maintain a drug concentration above the threshold concentration required to induce weight loss for a prolonged period.

In another embodiment, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to a prediabetic subject as an oral, transdermal or intranasal formulation to reach and maintain a drug concentration above the threshold concentration required to prevent the transition to diabetes for a prolonged period.

In another embodiment, a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to a type 1 diabetic subject as an oral, transdermal or intranasal formulation to reach and maintain a drug concentration above the threshold concentration required to induce beta cell rest for a prolonged period.

In another embodiment, a single dose of a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an subject in need thereof that results in circulating concentration of active drug sufficient to diminish the secretion of insulin for 24 or more hours.

In another embodiment, a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered over a prolonged basis to an subject in need thereof no more than once per 24 hours that results in circulating concentration of active drug sufficient to diminish the secretion of insulin on a continuous basis.

In another embodiment, a single dose of a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an subject in need thereof that results in circulating concentration of active drug sufficient to elevate non-esterified fatty acids in circulation for 24 or more hours.

In another embodiment, a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered over a prolonged basis to an subject in need thereof no more than once per 24 hours that results in circulating concentration of active drug sufficient to elevate non-esterified fatty acids in circulation on a continuous basis.

In another embodiment, a single dose of a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an subject in need thereof that results in circulating concentration of active drug sufficient to treat hypoglycemia in circulation for 24 or more hours.

In another embodiment, a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered over a prolonged basis to an subject in need thereof no more than once per 24 hours that results in circulating concentration of active drug sufficient to treat hypoglycemia on a continuous basis.

In another embodiment, a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered over a prolonged basis to an subject in need thereof no more than once per 24 hours that results in circulating concentration of active drug sufficient to induce weight loss on a continuous basis.

In another embodiment, a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered over a prolonged basis to an subject in need thereof no more than once per 24 hours that results in circulating concentration of active drug sufficient to maintain weight on a continuous basis, as it is preferable to maintain weight in an obese subject once some weight loss has occurred when the alternative is to regain weight.

In another embodiment, a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered over a prolonged basis to an subject in need thereof no more than once per 24 hours that results in circulating concentration of active drug sufficient to reduce circulating triglyceride levels on a continuous basis.

In another embodiment, a single dose of a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered to an subject in need thereof that results in circulating concentration of active drug sufficient to reduce or prevent ischemic or reperfusion injury in circulation for 24 or more hours.

In another embodiment, a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is administered over a prolonged basis to an subject in need thereof no more than once per 24 hours that results in circulating concentration of active drug sufficient reduce or prevent ischemic or reperfusion injury on a continuous basis.

In another embodiment, the frequency of adverse effects caused by treatment with a KATP channel opener selected from a salt of a compound of Formulae I-VIII is reduced using a pharmaceutically acceptable formulation of diazoxide or its derivatives that is administered to an subject in need thereof on a daily basis in which the first dose is known to be subtherapeutic and daily dose is subsequently increased stepwise until the therapeutic dose is reached.

In another embodiment, the frequency of adverse effects caused by treatment with a KATP channel opener selected from a salt of a compound of Formulae I-VIII is reduced using a pharmaceutically acceptable formulation that is administered to an subject in need thereof on a daily basis in which the active ingredient is not released from the formulation until gastric transit is complete.

In another embodiment, the frequency of adverse effects caused by treatment with a KATP channel opener selected from a salt of a compound of Formulae I-VIII is reduced using a pharmaceutically acceptable formulation that is administered to an subject in need thereof on a daily basis in which the maximum circulating concentration of active ingredient is lower than what would be realized by the administration of the same dose using an oral suspension or capsule formulation of Proglycem®.

In another embodiment, the frequency of adverse effects caused by treatment with a KATP channel opener selected from a salt of a compound of Formulae I-VIII is reduced using a pharmaceutically acceptable formulation that is administered to an subject in need thereof on a daily basis in which the first dose is known to be subtherapeutic and daily dose is subsequently increased stepwise until the therapeutic dose is reached, the active ingredient is not release from the formulation until gastric transit is complete and in which the maximum circulating concentration of active ingredient is lower than what would be realized by the administration of the same dose using an oral suspension or capsule formulation of Proglycem®.

In another embodiment, the frequency of adverse effects caused by treatment with a KATP channel opener selected from a salt of a compound of Formulae I-VIII is reduced using a pharmaceutically acceptable formulation that is administered to an overweight or obese subject in need thereof on a daily basis in which the first dose is known to be subtherapeutic and daily dose is subsequently increased stepwise until the therapeutic dose is reached, the active ingredient is not release from the formulation until gastric transit is complete, in which the maximum circulating concentration of active ingredient is lower than what would be realized by the administration of the same dose using an oral suspension or capsule formulation of Proglycem®, and in which the maximum dose is less than 5 mg/kg/day.

In another embodiment, the frequency of adverse effects caused by treatment with a KATP channel opener selected from a salt of a compound of Formulae I-VIII is reduced using a pharmaceutically acceptable formulation that is administered to an overweight or obese subject in need thereof on a daily basis in which the first dose is known to be subtherapeutic and daily dose is subsequently increased stepwise until the therapeutic dose is reached, the active ingredient is not release from the formulation until gastric transit is complete, in which the maximum circulating concentration of active ingredient is lower than what would be realized by the administration of the same dose using an oral suspension or capsule formulation, and in which the maximum dose is less than 2.5 mg/kg/day.

In another embodiment, the treatment of an overweight or obese subject is optimized for weight loss by administration of a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII once per 24 hours in which the release of the active ingredient from the formulation has been modified to provide continuous release for at least 6 hours.

In another embodiment, the treatment of an overweight or obese subject is optimized for weight loss by administration of a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII once per 24 hours in which the release of the active ingredient from the formulation has been modified to provide continuous release for at least 12 hours.

In another embodiment, the treatment of an overweight or obese subject is optimized for weight loss by administration of a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII once per 24 hours in which the release of the active ingredient from the formulation has been modified to provide a rising drug concentration in circulation for at least 8 hours.

In another embodiment, the treatment of an overweight or obese subject is optimized for weight loss by administration of a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII once per 24 hours in which the release of the active ingredient from the formulation has been modified to provide a rising drug concentration in circulation for at least 12 hours.

In another embodiment, the treatment of an overweight or obese subject is optimized for weight loss by administration of a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII once per 24 hours in which the release of the active ingredient from the formulation has been modified to match the pattern of basal insulin secretion.

In another embodiment, the frequency of adverse effects caused by treatment with a KATP channel opener selected from a salt of a compound of Formulae I-VIII is reduced using a pharmaceutically acceptable formulation that is administered to an obesity prone subject in need thereof on a daily basis in which the first dose is known to be subtherapeutic and daily dose is subsequently increased stepwise until the therapeutic dose is reached, the active ingredient is not release from the formulation until gastric transit is complete, in which the maximum circulating concentration of active ingredient is lower than what would be realized by the administration of the same dose using an oral suspension or capsule formulation, and in which the maximum dose is less than 5 mg/kg/day.

In another embodiment, the frequency of adverse effects caused by treatment with a KATP channel opener selected from a salt of a compound of Formulae I-VIII is reduced using a pharmaceutically acceptable formulation that is administered to an obesity prone subject in need thereof on a daily basis in which the first dose is known to be subtherapeutic and daily dose is subsequently increased stepwise until the therapeutic dose is reached, the active ingredient is not release from the formulation until gastric transit is complete, in which the maximum circulating concentration of active ingredient is lower than what would be realized by the administration of the same dose using an oral suspension or capsule formulation, and in which the maximum dose is less than 2.5 mg/kg/day.

In another embodiment, the treatment of an obesity prone subject is optimized for weight maintenance by administration of a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII once per 24 hours in which the release of the active ingredient from the formulation has been modified to provide continuous release for at least 6 hours.

In another embodiment, the treatment of an obesity prone subject is optimized for weight maintenance by administration of a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII once per 24 hours in which the release of the active ingredient from the formulation has been modified to provide continuous release for at least 12 hours.

In another embodiment, the treatment of an obesity prone subject is optimized for weight maintenance by administration of a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII once per 24 hours in which the release of the active ingredient from the formulation has been modified to provide a rising drug concentration in circulation for at least 8 hours.

In another embodiment, the treatment of an obesity prone subject is optimized for weight maintenance by administration of a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII once per 24 hours in which the release of the active ingredient from the formulation has been modified to provide a rising drug concentration in circulation for at least 12 hours.

In another embodiment, the treatment of an obesity prone subject is optimized for weight maintenance by administration of a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII once per 24 hours in which the release of the active ingredient from the formulation has been modified to match the pattern of basal insulin secretion.

In another embodiment, a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is co-administered with sibutramine to an overweight or obese subject to induce weight loss.

In another embodiment, a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is co-administered with orlistat to an overweight or obese subject to induce weight loss.

In another embodiment, a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is co-administered with rimonabant to an overweight or obese subject to induce weight loss.

In another embodiment, a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is co-administered with an appetite suppressant to an overweight or obese subject to induce weight loss.

In another embodiment, a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is co-administered with an anti-depressant to an overweight or obese subject to induce weight loss.

In another embodiment, a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is co-administered with anti-epileptic to an overweight or obese subject to induce weight loss.

In another embodiment, a pharmaceutically acceptable formulation of a KATP channel opener is selected from a salt of a compound of Formulae I-VIII is co-administered with a non-thiazide diuretic to an overweight or obese subject to induce weight loss.

In another embodiment, a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is co-administered with a drug that induces weight loss by a mechanism that is distinct from diazoxide to an overweight or obese subject to induce weight loss.

In another embodiment, a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is co-administered with a drug that lowers blood pressure to an overweight, obesity prone or obese subject to induce weight loss and treat obesity associated co-morbidities.

In another embodiment, a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is co-administered with a drug that lowers cholesterol to an overweight, obesity prone or obese subject to induce weight loss and treat obesity associated co-morbidities.

In another embodiment, a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is co-administered with a drug that raises HDL associated cholesterol to an overweight, obesity prone or obese subject to induce weight loss and treat obesity associated co-morbidities.

In another embodiment, a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is co-administered with a drug that improves insulin sensitivity to an overweight, obesity prone or obese subject to induce weight loss and treat obesity associated co-morbidities.

In another embodiment, a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is co-administered with a an anti-inflammatory to an overweight, obesity prone or obese subject to induce weight loss and treat obesity associated co-morbidities.

In another embodiment, a pharmaceutically acceptable formulation of a KATP channel opener selected from a salt of a compound of Formulae I-VIII is co-administered with a drug that lowers circulating triglycerides to an overweight, obesity prone or obese subject to induce weight loss and treat obesity associated co-morbidities.

In another embodiment, KATP channel openers selected from salts of compounds of Formulae I-VIII are co-formulated with sibutramine in a pharmaceutically acceptable formulation that is administered to an overweight, obesity prone or obese subject to induce weight loss and treat obesity-associated co-morbidities.

In another embodiment, KATP channel openers selected from salts of compounds of Formulae I-VIII are co-formulated with orlistat or other active that suppresses the action of gastric lipases in a pharmaceutically acceptable formulation that is administered to an overweight, obesity prone or obese subject to induce weight loss and treat obesity-associated co-morbidities.

In another embodiment, KATP channel openers selected from salts of compounds of Formulae I-VIII are co-formulated with a non-thiazide diuretic in a pharmaceutically acceptable formulation that is administered to an overweight, obesity prone or obese subject to induce weight loss and treat obesity-associated co-morbidities.

In another embodiment, KATP channel openers selected from salts of compounds of Formulae I-VIII are co-formulated with an appetite suppressant in a pharmaceutically acceptable formulation that is administered to an overweight, obesity prone or obese subject to induce weight loss and treat obesity-associated co-morbidities.

In another embodiment, KATP channel openers selected from salts of compounds of Formulae I-VIII are co-formulated with a cannabinoid receptor antagonist in a pharmaceutically acceptable formulation that is administered to an overweight, obesity prone or obese subject to induce weight loss and treat obesity-associated co-morbidities.

In another embodiment, KATP channel openers selected from salts of compounds of Formulae I-VIII are co-formulated with an anti-cholesteremic active in a pharmaceutically acceptable formulation that is administered to an overweight, obesity prone or obese subject to induce weight loss and treat obesity-associated co-morbidities.

In another embodiment, KATP channel openers selected from salts of compounds of Formulae I-VIII are co-formulated with an antihypertensive active in a pharmaceutically acceptable formulation that is administered to an overweight, obesity prone or obese subject to induce weight loss and treat obesity-associated co-morbidities.

In another embodiment, KATP channel openers selected from salts of compounds of Formulae I-VIII are co-formulated with an insulin sensitizing active in a pharmaceutically acceptable formulation that is administered to an overweight, obesity prone or obese subject to induce weight loss and treat obesity-associated co-morbidities.

In another embodiment, KATP channel openers selected from salts of compounds of Formulae I-VIII are co-formulated with an anti-inflammatory active in a pharmaceutically acceptable formulation that is administered to an overweight, obesity prone or obese subject to induce weight loss and treat obesity-associated co-morbidities.

In another embodiment, KATP channel openers selected from salts of compounds of Formulae I-VIII are co-formulated with an anti-depressant active in a pharmaceutically acceptable formulation that is administered to an overweight, obesity prone or obese subject to induce weight loss and treat obesity-associated co-morbidities.

In another embodiment, KATP channel openers selected from salts of compounds of Formulae I-VIII are co-formulated with an anti-epileptic active in a pharmaceutically acceptable formulation that is administered to an overweight, obesity prone or obese subject to induce weight loss and treat obesity-associated co-morbidities.

In another embodiment, KATP channel openers selected from salts of compounds of Formulae I-VIII are co-formulated with an active that reduces the incidence of atherosclerotic plaque in a pharmaceutically acceptable formulation that is administered to an overweight, obesity prone or obese subject to induce weight loss and treat obesity-associated co-morbidities.

In another embodiment, KATP channel openers selected from salts of compounds of Formulae I-VIII are co-formulated with an active that lowers circulating concentrations of triglycerides in a pharmaceutically acceptable formulation that is administered to an overweight, obesity prone or obese subject to induce weight loss and treat obesity-associated co-morbidities.

The reduction of circulating triglycerides in an overweight, obese or obesity prone subject is achieved by the administration of an effective amount of an oral dosage form of a KATP channel opener. selected from a salt of a compound of Formulae I-VIII.

An oral dosage form of KATP channel opener selected from a salt of a compound of Formulae I-VIII can be used to administer a therapeutically effective dose of KATP channel opener to an overweight or obesity prone subject in need thereof to maintain weight, as it is preferable to maintain weight in an obese subject once some weight loss has occurred when the alternative is to regain weight.

In another embodiment of the invention, KATP channel openers selected from salts of compounds of Formulae I-VIII are co-formulated with a drug to treat obesity. Such co-formulations can be formulated for oral administration once per 24 hours, for delayed release of the active until gastric transit is complete, and for sustained release of the active over a period of 2 to 24 hours. Such obesity treatment drugs include, but are not limited to: sibutramine hydrochloride (5-30 mg), orlistat (50-360 mg), phentermine hydrochloride or resin complex (15 to 40 mg), zonisamide (100 to 600 mg), topiramate (64 to 400 mg), naltrexone hydrochloride (50 to 600 mg), or rimonabant (5 to 20 mg).

A further embodiment of the co-formulation contains KATP channel openers selected from salts of compounds of Formulae I-VIII and a drug to treat obesity. Such co-formulations can be formulated for oral administration twice per 24 hours, for delayed release of the active until gastric transit is complete, and for sustained release of the active over a period of 2 to 12 hours. Such obesity treatment drugs include, but are not limited to: sibutramine hydrochloride (2.5 to 15 mg), orlistat (25 to 180 mg), phentermine hydrochloride or resin complex (7.5 to 20 mg), zonisamide (50 to 300 mg), topiramate (32 to 200 mg), naltrexone hydrochloride (25 to 300 mg), or rimonabant (2.5 to 10 mg).

In another embodiment of the invention KATP channel openers selected from salts of compounds of Formulas I-VIII are co-formulated with a drug to treat obesity, diabetes, metabolic syndrome or an obesity related comorbidity. Such drugs to treat these conditions include drugs that: agonizes the α1-noradrenergic receptor; agonizes the β2 noradrenergic receptor; stimulates noradrenalin release; blocks noradrenalin uptake; stimulates 5-HT release; blocks 5-HT uptake; is a serotonin (5-hydroxytryptamine) 2C receptor agonist; antagonizes acetyl-CoA carboxylase 2; agonizes the D1-receptor; antagonizes the H3-receptor; is a leptin analogue; agonizes the leptin receptor, sensitizes CNS tissue to the action of leptin; agonizes the MC4 receptor; agonizes NPY-Y1; agonizes NPY-Y2; agonizes NPY-Y4; agonizes NPY-Y5; antagonizes the MCH receptor; blocks CRH-BP; agonizes the CRH receptor; agonizes the urocortin receptor; antagonizes the galanin receptor; antagonizes the orexin receptor; agonizes the CART receptor; agonizes the Amylin receptor; agonizes the Apo(AIV) receptor; antagonizes the CB-1 receptor; is an αMSH analogue; inhibits PTP-1B; antagonizes PPARγ receptor; is a short acting bromocriptine; agonizes somatostatin; increases adiponectin; increases CCK activity; increases PYY activity; increases GLP-1 activity; decreases ghrelin activity; is a selective B3 stimulator or agonist; agonizes thyroid receptor; inhibits gastrointestinal lipases or other digestive enzymes; blocks absorption of dietary fat; or block de-novo fatty acid synthesis. Additionally, such drugs to treat obesity may include, but are not limited to those that antagonize or agonize the function or expression of 11B hydroxysteroid dehydrogenase type 1; acetyl-CoA carboxylase 1; ADAM 12, member 12 of a disintegrin and metalloprotease family or its shorter secreted form; agouti related protein; angiotensinogen; adipocyte lipid binding protein; adipocyte fatty acid binding protein; adrenergic receptors; acylation-stimulating protein; bombesin receptor subtype-3; C/EBP, CCAAT/enhancer binding protein; cocaine- and amphetamine-regulated transcript; cholecystokinin; cholecystokinin A receptor; CD36, fatty acid translocase; corticotropin-releasing hormone; diacylglycerol acyltransferases; E2F transcription factor; eukaryotic translation initiation factor 4e binding protein 1; estrogen receptor; fatty acid synthase; fibroblast growth factor; forkhead box C2; glucose-dependent insulinotropic peptide; GIP receptor; inhibitory G protein alpha-subunit; glucagon-like peptide-1; GLP-1 receptor; glycerol-3-phosphate acyltransferase; glycerol 3-phosphate dehydrogenase; stimulatory G protein alpha-subunit; high-mobility group phosphoprotein isoform I-C; hormone sensitive lipase; inducible nitric oxide synthase; Janus kinases; lipoprotein lipase; melanocortin-3 receptor; melanocortin-4 receptor; mitochondrial GPAT; metallothionein-I and -II; nescient helix-loop-helix 2; neuropeptide Y; neuropeptide Y-1 receptor; neuropeptide Y-2 receptor; neuropeptide Y-4 receptor; neuropeptide Y-5 receptor; plasminogen activator inhibitor-1; PPARgamma co-activator 1; pro-opiomelanocortin; peroxisome proliferator-activated receptor; protein tyrosine phosphatase 1B; regulatory subunit IIbeta of protein kinase A; retinoid X receptor; steroidogenic factor 1; single-minded 1; sterol regulatory element binding protein; tyrosine hydroxylase; thyroid hormone receptor a2; uncoupling protein; nerve growth factor induced protein; leucine zipper transcription factor; a-melanocyte-stimulating hormone.

In another embodiment of the invention, KATP channel openers selected from salts of compounds of Formulae I-VIII are co-formulated with a drug to treat diabetes. Such co-formulations can be formulated for oral administration once per 24 hours, for delayed release of the active until gastric transit is complete, and for sustained release of the active over a period of 2 to 24 hours. Such diabetes treatment drugs include, but are not limited to: acarbose (50 to 300 mg), miglitol (25 to 300 mg), metformin hydrochloride (300 to 2000 mg), repaglinide (1-16 mg), nateglinide (200 to 400 mg), or rosiglitazone (5 to 50 mg).

In a further embodiment, the co-formulation can be formulated for oral administration twice per 24 hours, for delayed release of the active until gastric transit is complete, and for sustained release of the active over a period of 2 to 12 hours. Such drugs to treat diabetes include, but are not limited to: acarbose (25 to 150 mg), miglitol (12.5 to 150 mg), metformin hydrochloride (150 to 1000 mg), repaglinide (0.5 to 8 mg), nateglinide (100 to 200 mg), or rosiglitazone (2.5 to 25 mg).

In another embodiment of the invention, KATP channel openers selected from salts of compounds of Formulae I-VIII are co-formulated with a drug to treat elevated cholesterol. Such co-formulations can be formulated for oral administration once per 24 hours, for delayed release of the active until gastric transit is complete, and for sustained release of the active over a period of 2 to 24 hours. Such drugs to treat elevated cholesterol include, but are not limited to: pravastatin, simvastatin, atorvastatin, fluvastatin, rosuvastatin or lovastatin (10 to 80 mg).

In a further embodiment, the co-formulation can be formulated for oral administration twice per 24 hours, for delayed release of the active until gastric transit is complete, and for sustained release of the active over a period of 2 to 12 hours. Such drugs to treat elevated cholesterol include, but are not limited to: pravastatin, simvastatin, atorvastatin, fluvastatin, rosuvastatin or lovastatin (5 to 40 mg).

In another embodiment of the invention, KATP channel openers selected from salts of compounds of Formulae I-VIII are co-formulated with a drug to treat depression. Such co-formulations can be formulated for oral administration once per 24 hours, for delayed release of the active until gastric transit is complete, and for sustained release of the active over a period of 2 to 24 hours. Such drugs to treat depression include, but are not limited to: citalopram hydrobromide (10 to 80 mg), escitalopram hydrobromide (5 to 40 mg), fluvoxamine maleate (25 to 300 mg), paroxetine hydrochloride (12.5 to 75 mg), fluoxetine hydrochloride (30 to 100 mg), setraline hydrochloride (25 to 200 mg), amitriptyline hydrochloride (10 to 200 mg), desipramine hydrochloride (10 to 300 mg), nortriptyline hydrochloride (10 to 150 mg), duloxetine hydrochloride (20 to 210 mg), venlafaxine hydrochloride (37.5 to 150 mg), phenelzine sulfate (10 to 30 mg), bupropion hydrochloride (200 to 400 mg), or mirtazapine (7.5 to 90 mg).

In a further embodiment, the co-formulation can be formulated for oral administration twice per 24 hours, for delayed release of the active until gastric transit is complete, and for sustained release of the active for 2 to 12 hours. Such drugs to treat depression include, but are not limited to: citalopram hydrobromide (5 to 40 mg), escitalopram hydrobromide (2.5 to 20 mg), fluvoxamine maleate (12.5 to 150 mg), paroxetine hydrochloride (6.25 to 37.5 mg), fluoxetine hydrochloride (15 to 50 mg), setraline hydrochloride (12.5 to 100 mg), amitriptyline hydrochloride (5 to 100 mg), desipramine hydrochloride (5 to 150 mg), nortriptyline hydrochloride (5 to 75 mg), duloxetine hydrochloride (10 to 100 mg), venlafaxine hydrochloride (18 to 75 mg), phenelzine sulfate (5 to 15 mg), bupropion hydrochloride (100 to 200 mg), or mirtazapine (4 to 45 mg).

In another embodiment of the invention, KATP channel openers selected from salts of compounds of Formulae I-VIII are co-formulated with a drug to treat hypertension. Such co-formulations can be formulated for oral administration once per 24 hours, for delayed release of the active until gastric transit is complete, and for sustained release of the active over a period of 2 to 24 hours. Such drugs to treat hypertension include, but are not limited to: enalapril maleate (2.5 to 40 mg), captopril (2.5 to 150 mg), lisinopril (10 to 40 mg), benzaepril hydrochloride (10 to 80 mg), quinapril hydrochloride (10 to 80 mg), peridopril erbumine (4 to 8 mg), ramipril (1.25 to 20 mg), trandolapril (1 to 8 mg), fosinopril sodium (10 to 80 mg), moexipril hydrochloride (5 to 20 mg), losartan potassium (25 to 200 mg), irbesartan (75 to 600 mg), valsartan (40 to 600 mg), candesartan cilexetil (4 to 64 mg), olmesartan medoxamil (5 to 80 mg), telmisartan (20 to 160 mg), eprosartan mesylate (75 to 600 mg), atenolol (25 to 200 mg), propranolol hydrochloride (10 to 180 mg), metoprolol tartrate, succinate or fumarate (25 to 400 mg), nadolol (20 to 160 mg), betaxolol hydrochloride (10 to 40 mg), acebutolol hydrochloride (200 to 800 mg), pindolol (5 to 20 mg), bisoprolol fumarate (5 to 20 mg), nifedipine (15 to 100 mg), felodipine (2.5 to 20 mg), amlodipine besylate (2.5 to 20 mg), nicardipine (10 to 40 mg), nisoldipine (10 to 80 mg), terazosin hydrochloride (1 to 20 mg), doxaxosin mesylate (4 to 16 mg), prazosin hydrochloride (2.5 to 10 mg), or alfuzosin hydrochloride (10 to 20 mg).

In a further embodiment, the co-formulation can be formulated for oral administration twice per 24 hours, for delayed release of the active until gastric transit is complete, and for sustained release of active over a period of 2 to 12 hours. Such drugs to treat hypertension include, but are not limited to: enalapril maleate (1.25 to 20 mg), captopril (2 to 75 mg), lisinopril (5 to 20 mg), benzaepril hydrochloride (5 to 40 mg), quinapril hydrochloride (5 to 40 mg), peridopril erbumine (2 to 4 mg), ramipril (1 to 10 mg), trandolapril (1 to 4 mg), fosinopril sodium (5 to 40 mg), moexipril hydrochloride (2.5 to 10 mg), losartan potassium (12.5 to 100 mg), irbesartan (37.5 to 300 mg), valsartan (20 to 300 mg), candesartan cilexetil (2 to 32 mg), olmesartan medoxamil (2.5 to 40 mg), telmisartan (10 to 80 mg), eprosartan mesylate (37.5 to 300 mg), atenolol (12.5 to 100 mg), propranolol hydrochloride (5 to 90 mg), metoprolol tartrate, succinate or fumarate (12.5 to 200 mg), nadolol (10 to 80 mg), betaxolol hydrochloride (5 to 20 mg), acebutolol hydrochloride (100 to 400 mg), pindolol (2.5 to 10 mg), bisoprolol fumarate (2.5 to 10 mg), nifedipine (7.5 to 50 mg), felodipine (1 to 10 mg), amlodipine besylate (1 to 10 mg), nicardipine (5 to 20 mg), nisoldipine (5 to 40 mg), terazosin hydrochloride (1 to 10 mg), doxaxosin mesylate (2 to 8 mg), prazosin hydrochloride (1 to 5 mg), or alfuzosin hydrochloride (5 to 10 mg).

In another embodiment of the invention, KATP channel openers selected from salts of compounds of Formulae I-VIII are co-formulated with a diuretic. Such co-formulations can be formulated for oral administration once per 24 hours, for delayed release of the active until gastric transit is complete, and for sustained release of the active over a period of 2 to 24 hours. Such diuretics can include, but are not limited to: amiloride hydrochloride (1 to 10 mg), spironolactone (10 to 100 mg), triamterene (25 to 200 mg), bumetanide (0.5 to 4 mg), furosemide (10 to 160 mg), ethacrynic acid or ethacrynate sodium (10 to 50 mg), torsemide (5 to 100 mg), chlorthalidone (10 to 200 mg), indapamide (1 to 5 mg), hydrochlorothiazide (10 to 100 mg), chlorothiazide (50 to 500 mg), bendroflumethiazide (5 to 25 mg), hydroflumethiazide (10 to 50 mg), mythyclothiazide (1 to 5 mg), and polythiazide (1 to 10 mg).

In a further embodiment, the co-formulation can be formulated for oral administration twice per 24 hours, for delayed release of the active until gastric transit is complete, and for sustained release of the active over a period of 2 to 12 hours. Such diuretics include, but are not limited to: amiloride hydrochloride (0.5 to 5 mg), spironolactone (5 to 50 mg), triamterene (12 to 100 mg), bumetanide (0.2 to 2 mg), furosemide (5 to 80 mg), ethacrynic acid or ethacrynate sodium (5 to 25 mg), torsemide (2 to 50 mg), chlorthalidone (5 to 100 mg), indapamide (0.5 to 2.5 mg), hydrochlorothiazide (5 to 50 mg), chlorothiazide (25 to 250 mg), bendroflumethiazide (2 to 12.5 mg), hydroflumethiazide (5 to 25 mg), mythyclothiazide (0.5 to 2.5 mg), and polythiazide (0.5 to 5 mg).

In another embodiment of the invention, KATP channel openers selected from salts of compounds of Formulae I-VIII are co-formulated with a drug to treat inflammation or pain. Such co-formulations can be formulated for oral administration once per 24 hours, for delayed release of the active until gastric transit is complete, and for sustained release of the active over a period of 2 to 24 hours. Such drugs to treat inflammation or pain include, but are not limited to: aspirin (100 to 1000 mg), tramadol hydrochloride (25 to 150 mg), gabapentin (100 to 800 mg), acetaminophen (100 to 1000 mg), carbamazepine (100 to 400 mg), ibuprofen (100 to 1600 mg), ketoprofen (12 to 200 mg), fenprofen sodium (100 to 600 mg), flurbiprofen sodium or flurbiprofen (50 to 200 mg), or combinations of these with a steroid or aspirin.

In a further embodiment, the co-formulation can be formulated for oral administration twice per 24 hours, for delayed release of the active until gastric transit is complete, and for sustained release of the active over a period of 2 to 12 hours. Such drugs to treat inflammation or pain include, but are not limited to: aspirin (100 to 650 mg), tramadol hydrochloride (12 to 75 mg), gabapentin (50 to 400 mg), acetaminophen (50 to 500 mg), carbamazepine (50 to 200 mg), ibuprofen (50 to 800 mg), ketoprofen (6 to 100 mg), fenprofen sodium (50 to 300 mg), flurbiprofen sodium or flurbiprofen (25 to 100 mg), or combinations of these with a steroid or aspirin.

A method of inducing loss of greater than 25% of initial body fat in an overweight or obese subject can be achieved by the prolonged administration of an oral dosage form of a KATP channel opener. selected from a salt of a compound of Formulae I-VIII.

A method of inducing loss of greater than 50% of initial body fat in an overweight or obese subject can be achieved by the prolonged administration of an oral dosage form of a KATP channel opener selected from a salt of a compound of Formulae I-VIII.

A method of inducing loss of greater than 75% of initial body fat in an overweight or obese subject can be achieved by the prolonged administration of an oral dosage form of a KATP channel opener. selected from a salt of a compound of Formulae I-VIII.

A method of inducing preferential loss of visceral fat in an overweight or obese subject can be achieved by the prolonged administration of an oral dosage form of a KATP channel opener. selected from a salt of a compound of Formulae I-VIII.

A method of inducing loss of body fat and reductions in circulating triglycerides in an overweight or obese subject can be achieved by the prolonged administration of an oral dosage form of a KATP channel opener. selected from a salt of a compound of Formulae I-VIII.

In some embodiments, the invention provides a polymorph of a salt, which salt includes diazoxide and a cation selected from the group consisting of an alkali metal and a compound comprising a tertiary amine or quaternary ammonium group. In some embodiments, the cation is choline.

In some embodiments, the polymorph of diazoxide choline salt is of Form A having characteristic peaks in the XRPD pattern at values of two-theta (Cu K, 40 kV, 40 mA) at approximately 9.8, 10.5, 14.9, 17.8, 17.9, 18.5, 19.5, 22.1, 22.6, 26.2, 29.6, and 31.2 degrees.

In some embodiments, the polymorph of diazoxide choline salt is of Form B having characteristic peaks in the XRPD pattern at values of two-theta (Cu K, 40 kV, 40 mA) at approximately 8.9, 10.3, 12.0, 18.3, 20.6, 24.1, 24.5, 26.3, 27.1, and 28.9 degrees.

In some embodiments, the polymorph of diazoxide choline salt is of Form A having characteristic infrared absorbances at 2926, 2654, 1592, 1449, and 1248 cm-1.

In some embodiments, the polymorph of diazoxide choline salt is of Form B having characteristic infrared absorbances at 3256, 2174, 2890, 1605, 1463, and 1235 cm-1.

In some embodiments, the polymorph of diazoxide includes potassium as the cation.

In some embodiments, the polymorph of diazoxide potassium salt is of Form A having characteristic peaks in the XRPD pattern at values of two-theta (Cu K, 40 kV, 40 mA) at approximately 6.0, 8.1, 16.3, 17.7, 18.6, 19.1, 22.9, 23.3, 23.7, 24.7, 25.4, 26.1, 28.2, 29.6, and 30.2 degrees.

In some embodiments, the polymorph of diazoxide potassium salt is of Form B having characteristic peaks in the XRPD pattern at values of two-theta (Cu K, 40 kV, 40 mA) at approximately 8.5, 10.8, 16.9, 18.2, 21.6, 25.5, 26.1, and 28.9 degrees.

In some embodiments, the polymorph of diazoxide potassium salt is of Form C having characteristic peaks in the XRPD pattern at values of two-theta (Cu K, 40 kV, 40 mA) at approximately 5.7, 6.1, 17.9, 23.9, 25.1, and 37.3 degrees.

In some embodiments, the polymorph of diazoxide potassium salt is of Form D having characteristic peaks in the XRPD pattern at values of two-theta (Cu K, 40 kV, 40 mA) at approximately 5.7, 6.2, 8.1, 8.5, 8.8, 16.9, 18.6, 23.2, 24.5, 25.8, and 26.1 degrees.

In some embodiments, the polymorph of diazoxide potassium salt is of Form E having characteristic peaks in the XRPD pattern at values of two-theta (Cu K, 40 kV, 40 mA) at approximately 6.7, 7.1, 14.1, and 21.2 degrees.

In some embodiments, the polymorph of diazoxide potassium salt is of Form F having characteristic peaks in the XRPD pattern at values of two-theta (Cu K, 40 kV, 40 mA) at approximately 8.5, 9.0, 18.7, 20.6, 23.5, 27.5, and 36.3 degrees.

In some embodiments, the polymorph of diazoxide potassium salt is of Form G having characteristic peaks in the XRPD pattern at values of two-theta (Cu K, 40 kV, 40 mA) at approximately 5.2, 5.5, 13.1, 16.5, 19.3, 22.8, 24.8, 26.4, 28.7, and 34.1 degrees.

In some embodiments, the polymorph of diazoxide potassium salt is of Form A having characteristic infrared absorbances at 1503, 1374, 1339, 1207, 1131, 1056, and 771 cm-1.

In some embodiments, the polymorph of diazoxide potassium salt is of Form B having characteristic infrared absorbances at 1509, 1464, 1378, and 1347 cm-1.

In some embodiments, the polymorph of diazoxide potassium salt is of Form C having characteristic infrared absorbances at 1706, 1208, 1146, and 746 cm-1.

In some embodiments, the polymorph of diazoxide potassium salt is of Form D having characteristic infrared absorbances at 1595, 1258, 1219, and 890 cm-1.

In some embodiments, the polymorph of diazoxide potassium salt is of Form E having characteristic infrared absorbances at 1550, 1508, 1268, 1101, and 1006 cm-1.

In some embodiments, the polymorph of diazoxide potassium salt is of Form F having characteristic infrared absorbances at 1643, 1595, 1234, 1145, and 810 cm-1.

In some embodiments, the polymorph of diazoxide potassium salt is of Form G having characteristic infrared absorbances at 1675, 1591, 1504, 1458, 1432, 1266, 999, 958, 905, and 872 cm-1.

In some embodiments, the polymorph of diazoxide choline salt is of Form A having characteristic peaks in the XRPD pattern substantially as shown in FIG. 16( a), and an NMR spectrum substantially as shown in FIG. 17( a).

In some embodiments, the polymorph of diazoxide choline salt is of Form B having characteristic peaks in the XRPD pattern substantially as shown in FIG. 16( c) and an NMR spectrum substantially as shown in FIG. 17( b).

In some embodiments, the polymorph of diazoxide potassium salt includes one or more of Forms A-G, wherein each of the Forms A-G has characteristic peaks in the XRPD pattern substantially as shown in FIG. 18-19.

In some embodiments of the invention there are provided methods for producing a diazoxide choline salt, which methods include suspending diazoxide in a solvent (e.g., alcohols such as methanol, i-BuOH, i-AmOH, t-BuOH, and the like, ketones, tetrahydrofuran, dimethylformamide, n-methylpyrrolidinone, and the like) and mixing with a choline salt (e.g., choline hydroxide), adding a co-solvent (e.g., MTBE, EtOA, IPA, c-Hexane, heptane, toluene, CH2CL2, dioxane, and the like) to the suspension under conditions sufficient to cause formation and precipitation of said diazoxide choline salt, and harvesting the precipitate to provide the diazoxide choline salt.

In some embodiments, the solvent is tetrahydrofuran. In some embodiments, the solvent is 2-methyltetrahydrofuran (2-MeTHF).

In some embodiments, the diazoxide and the solvent are present at a ratio of about 1 g diazoxide per 1 mL solvent to about 1 g diazoxide per 5 mL solvent. In some embodiments, the diazoxide and the solvent are present at a ratio of about 1 g diazoxide per 3 mL solvent.

In some embodiments, the choline salt is a solution in MeOH. In some embodiments, the choline salt is choline hydroxide in about a 45% solution (e.g., 40%, 41%, 42%, 43%, 44%, 45%, 46%,47%, 48%, 49%, 50%) in MeOH.

In some embodiments, the choline salt is added as 1 equivalent of diazoxide.

In some embodiments, the co-solvent is MTBE.

In some embodiments, the amount of co-solvent added is in a ratio to the amount of the solvent of about 3:14 (solvent:co-solvent) (e.g., 3:12, 3:13, 3:14, 3:15, 3:16).

In some embodiments, the process of making polymorphs of diazoxide choline salt includes the step of seeding with crystals of diazoxide choline salt polymorph Form B prior to the harvesting step.

In some embodiments for the method for producing a diazoxide choline salt the salt includes polymorph Form B substantially free of polymorph Form A, the polymorph Form B having characteristic peaks in the XRPD pattern at values of two-theta (Cu K, 40 kV, 40 mA) at approximately 8.9, 10.3, 12.0, 18.3, 20.6, 24.1, 24.5, 26.3, 27.1, and 28.9 degrees.

In some embodiments for the method of treating obesity or obesity-related morbidity in an obese subject, the compound is a compound of Formula V.

In some embodiments for the method of treating obesity or obesity-related morbidity in an obese subject, the compound is a compound of Formula VI.

In some embodiments for the method of treating obesity or obesity-related morbidity in an obese subject, the compound is a compound of Formula VII.

In some embodiments for the method of treating obesity or obesity-related morbidity in an obese subject, the compound is a compound of Formula VIII.

In some embodiments for the method of treating obesity or obesity-related morbidity in an obese subject, the method further comprises administering a drug selected from the group consisting of Sibutramine, Orlistat, Rimonabant, an appetite suppressant, a non-thiazide diuretic, a drug that lowers cholesterol, a drug that raises HDL cholesterol, a drug that lowers LDL cholesterol, a drug that lowers blood pressure, a drug that is an anti-depressant, a drug that is an anti-epileptic, a drug that is an anti-inflammatory, a drug that is an appetite suppressant, a drug that lowers circulating triglycerides, and a drug that is used to induce weight loss in an overweight or obese individual.

In some embodiments for the method of treating obesity or obesity-related morbidity in an obese subject, the method further comprises administering a pharmaceutically active agent other than the KATP channel opener. In some embodiments, the other pharmaceutically active agent is an agent useful for the treatment of a condition selected from the group consisting of obesity, prediabetes, diabetes, hypertension, depression, elevated cholesterol, fluid retention, obesity associated co-morbidities, ischemic and reperfusion injury, epilepsy, cognitive impairment, schizophrenia, mania, and other psychotic condition.

In some embodiments for the method for treating a subject suffering from or at risk for Alzheimer's disease (AD), the method includes administration to a subject a therapeutically effective amount of a salt of diazoxide including salts provided herein. In some embodiments for the method for treating a subject suffering from or at risk for AD, the method includes administration to a subject a therapeutically effective amount of a compound according to any of Formulae I-VIII. In some embodiments, the compound is diazoxide or a salt thereof.

AD is a neurodegenerative disorder neuropathologically characterized by abnormal accumulations of intracellular neurofibrilary tangles and extracellular amyloid plaques throughout cortical and limbic brain regions and the loss of synapses and neurons. AD is further characterized by significant cognitive and memory impairment. 13 amyloid plaques form from the β amyloid peptide, either 1-40 or 1-42 peptide, which is released from amyloid precursor protein following cleavage by gamma secretase. In addition to forming plaques, the β amyloid peptides are cytotoxic either as the monomer or as a short-lived oligomeric intermediate. β amyloid peptides (monomers, dimers or oligomers) can be identified both in CSF (cerebrospinal fluid) and in serum. Amyloid angiopathy is characterized by Aβ deposition and may contribute to the cerebrovascular abnormalities that precede the onset of AD.

In one embodiment, a therapeutically effective amount of a KATP channel opener, or pharmaceutical salt thereof, is formulated for once, twice or thrice per day administration for the treatment of patients diagnosed with hypercholesterolemia, combined hyperlipidemia, endogenous hyperlipidemia, or hypertriglyceridemia. These conditions include Fredrickson class IIa, IIb, IV and V (Fredrickson, D S, Lees, L S, Circulation 1965 31:321-327; Beaumont, J L et al. Bull World Health Org 1970 43(6):891-915).

Also provided are pharmaceutical coformulations comprising a therapeutically effective amount of a KATP channel opener, or pharmaceutical salt thereof, and one or more of the following:

a) an inhibitor of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase or pharmaceutical salt thereof;

b) a statin or pharmaceutical salt thereof;

c) 10 to 100 mg equivalent of atorvastatin, or a pharmaceutical salt thereof;

d) 10 mg to 100 mg of fluvastatin, or a pharmaceutical salt thereof;

e) 10 to 80 mg equivalent of lovastatin, or a pharmaceutical salt thereof;

f) 10 to 100 mg equivalent of mevastatin, or a pharmaceutical salt thereof;

g) pitavastatin, or a pharmaceutical salt thereof;

h) 10 to 100 mg equivalent of pravastatin, or a pharmaceutical salt thereof;

i) rosuvastatin, or a pharmaceutical salt thereof;

j) 5 to 100 mg equivalent of simvastatin, or a pharmaceutical salt thereof;

k) 5 to 20 mg equivalent of ezetimibe, or a pharmaceutical salt thereof;

l) a fibrate, or a pharmaceutical salt thereof;

m) 25 mg to 250 mg equivalent of fenofibrate, or a pharmaceutical salt thereof;

n) 200 mg to 600 mg equivalent of clofibrate, or a pharmaceutical salt thereof;

o) 200 mg to 700 mg equivalent of gemfibrozil, or a pharmaceutical salt thereof;

p) 100 mg to 800 mg equivalent of benzafibrate, or a pharmaceutical salt thereof;

q) 50 mg to 400 mg equivalent of ciprofibrate, or a pharmaceutical salt thereof;

r) 50 mg to 400 mg equivalent of ciprofibrate, or a pharmaceutical salt thereof;

s) histamine H3 antagonist, or a pharmaceutical salt thereof;

t) histamine H3 inverse agonist, or a pharmaceutical salt thereof;

u) absorbable or non-absorbable MTP inhibitor, or a pharmaceutical salt thereof;

v) a thyroid receptor activator, or a pharmaceutical salt thereof; or

w) a squalene synthase inhibitor, or a pharmaceutical salt thereof.

Any of the above pharmaceutical coformulations may be administered either once per day, twice per day or three times per day for the treatment of patients diagnosed with hypercholesterolemia, combined hyperlipidemia, endogenous hyperlipidemia, or hypertriglyceridemia. These conditions include Fredrickson class IIa, IIb, IV and V.

Treatment of Hypertriglyceridemic Patients with Omega-3 Fatty Acids and Diazoxide

Prescription omega-3 fatty acids (p-OM3) are often used to treat patients with elevated triglycerides, however fatty acid compositions differ among products, which can lead to different effects on levels of LDL-C in these patients. Some products contain very high levels of eicosapentaenoic acid (EPA) and are substantially free of docosahexaenoic acid (DHA). Exemplary products include AMR-101 (1 gram gel cap that contains at least 950 mg of ethyl EPA and is substantially free of the ethyl ester of DHA) and Epadel® (highly pure ethyl EPA packaged in gel caps dosages of 300 mg, 600 mg or 900 mg). Other products are not substantially free of DHA, such as Lovaza® or Omacor® (1 capsule contains about 465 mg of EPA and 375 mg of DHA).

Treatment of hypertriglyceridemic patients with a p-OM3 formulation high in EPA and substantially free of DHA generally results in a more modest improvement in triglycerides while causing a limited increase in LDL-C concentrations. In contrast, treatment with a p-OM3 formulation with EPA and higher amounts of DHA generally results in greater improvements in triglyceride levels but also can result in up to a nearly 50% increase in LDL-C. The increase in LDL-C results from the mode of action of omega 3 fatty acid products which include DHA, namely increasing clearance of triglycerides, while decreasing the size of secreted VLDL particles. Triglycerides synthesized by the liver circulate primarily as a component of apolipoprotein B-containing particles. These particles, which are atherogenic, contain both cholesterol and triglycerides, and are secreted as very low density lipoprotein particles. Triglycerides are removed from these particles by the action of lipoprotein lipase. As triglycerides are removed, the density of these particles gradually increases from very low density (VLDL) to intermediate density (IDL) and ultimately to low density (LDL). Drugs that reduce triglycerides by removal of triglycerides from Apo B-containing lipoprotein particles will cause the transition of VLDL to IDL and ultimately to LDL thereby raising the concentration of LDL-C in treated patients.

Thus, a very high triglyceride patient being treated with p-OM3 that contains DHA may be at increased risk for myocardial infarction or stroke because of elevated LDL-C. Treatment with p-OM3 that is substantially free of DHA can reduce this problem. Nevertheless, patients treated with a p-OM3 that is substantially free of DHA may fail to reach their triglyceride goal due to their moderate effects on reducing triglyceride levels. In these instances, such patients may benefit from co-administration of another triglyceride-lowering drug.

A preferred combination of drugs to treat dyslipidemia is one in which the mode of action of the drugs differs, and which offer substantial opportunity for complementary effects. However, some drug combinations do not provide an additive improvement in triglyceride levels.

For example, a clinical study has been conducted evaluating the value of co-administering the p-OM3 product (Lovaza®) with fenofibrate in the treatment of patients with very high triglycerides (Roth et al., J Cardiovasc Pharmacol. 2009 September; 54(3):196-203). The net result of addition of Lovaza® treatment for patients undergoing fenofibrate treatment for very high triglycerides was little further improvement in triglyceride levels compared to treatment with fenofibrate alone (see section 15.4 and FIG. 29). This lack of additional improvement is likely because both of drugs have similar modes of action, namely clearance of triglycerides from circulation. Furthermore, the combined use of Lovaza® with fenofibrate in the treatment of patients with very high triglycerides results in very marked increases in LDL cholesterol (see Example 15.4 and FIG. 29), which contributes to increased risk of major cardiovascular events.

For a patient with very high triglycerides who has elevated risk for major cardiovascular events attributable to elevated LDL-C, a more preferable drug combination for lowering triglycerides is a K_(ATP) channel agonist, e.g. diazoxide or a salt thereof, co-administered with a p-OM3 product (such as AMR-101 or Epadel®) that includes a high concentration of EPA and is substantially free of DHA. Treatment of very high triglyceride patients by co-administering a K_(ATP) channel agonist, e.g. diazoxide or a salt thereof, and a high EPA/low DHA p-OM3 product will result in improvements in triglycerides that are greater than those achieved with either product alone while at the same time minimizing the potential for increases in LDL cholesterol.

Treatment of Hypertriglyceridemic Patients by Coadministration of Fenofibrate and Diazoxide Choline

Fibrates, such as fenofibrate, are another class of drugs used to lower triglycerides, but alone may not be enough to allow hypertriglyceridemic patients to reach their triglyceride goal. As described above, fenofibrate co-administered with p-OM3 does not have a significant additive effect in reducing triglycerides, likely due to their similar modes of action (see also Example 15.4 and FIG. 29).

In newly diagnosed very high triglyceride patients who are not anticipated to reach their triglyceride goal with a single agent (i.e., typically patients with circulating triglycerides at diagnosis of 500 mg/dL or higher, such as 1000 mg/dL or higher), it may be preferable to initiate therapy with co-administration of a K_(ATP) channel agonist, e.g. diazoxide or a salt thereof, and a fibrate, such as fenofibrate or a salt thereof. In these combined therapies, the K_(ATP) channel agonist and the fibrate may be co-administered as part of separate formulations, or may be co-formulated.

There are a number of branded and generic fibrate products that differ in the extent of micronization of the fibrate, the excipients used in the formulation and the dosage form. The Trilipix® product, the leading branded fenofibrate product, makes use of fenofibrate choline as opposed to the fenofibric acid used in the other fenofibrate products. Most fenofibrate products are manufactured in two dose strengths, a starter or titration dose and a maintenance dose, which tends to contain approximately 3 times as much fenofibrate as the titration dose. Depending on the product, these doses may be within the ranges of 35 mg and 105 mg, and 40 mg and 120 mg, 45 mg and 135 mg, 48 mg and 145 mg, and 50 mg and 160 mg. Some fenofibrate products may be manufactured in 3 or more dose strengths. Depending on the product, these may include 50 mg, 100 mg and 150 mg; 43 mg, 87 mg and 130 mg; 45 mg, 90 mg, and 135 mg; or 67 mg, 100 mg, 134 mg, and 200 mg. Fibrates may be formulated as a capsule, tablet or delayed release capsule product.

Initiation of both a fibrate and a K_(ATP) channel agonist, e.g., diazoxide or a salt thereof, may be titrated; thus if treatment is initiated with both products simultaneously, the treated patient may receive the starter dose for both products. For example, a titration dose of a fibrate may be between about 35 to 100 mg, such as between about 35 mg to 67 mg; and a titration dose of a K_(ATP) channel agonist may be about 70 to 220 mg; such as between about 72.5 to 145 mg of a K_(ATP) channel agonist. Titration of fibrate, e.g. fenofibrate or a salt thereof, and a K_(ATP) channel agonist, e.g. diazoxide or a salt thereof, may extend over 7 to 28 days and may include at least one additional dose level prior to reaching the maintenance dose of either product. The maintenance dose on a fibrate may typically be about 2 times to 3 times the starter dose. For example, the maintenance dose may contain between 100 mg and 200 mg of fenofibrate. The maintenance dose of the K_(ATP) channel agonist, e.g. diazoxide or a salt thereof, may be about 2 times to 3 times the titration dose. For example a diazoxide choline maintenance dose may range from 145 to 435 mg.

To accommodate initiation of treatment with both products, a titration kit may be prepared in which each product is packaged for daily administration in a blister pack or similar arrangement. Each daily blister may contain either one dose of each drug product or the two drugs blistered separately. The blister material may be any standard blister material known in the art, such as plastic, foil, or a multi-layer product containing both aluminum and one or more layers of either PVC or nylon. Exemplary titration kits may contain from 7 blisters, where both drug products are in the same blister and titration extends only over 7 days, to more than 56 blisters, where each product is packaged separately and titration extends over 28 days. The kit may include up to two dose levels of each product and may also contain the maintenance dose for at least 1 day of treatment. Each blister may be identified as to the day of treatment, or the dose and day of treatment with that dose. The different dose strengths of each drug product may be uniquely identified as to dose either by color, combination of colors for the capsule product, markings or both color and markings. Alternatively, the blister cards may contain sufficient drug product for 7 days and the kit may contain one or more weekly cards. The cards may be identical or may differ and be identified as to dose strength of each product or week of treatment.

Intravenous Administration of Diazoxide and Insulin

The primary objectives of intervention in patients with acute pancreatitis (AP) who have elevated triglycerides are (1) to rapidly lower triglycerides (TG) and free fatty acids (FFA); (2) to limit pancreatic and peripancreatic necrosis; and (3) to limit organ failure and the associated mortality. The primary means of lowering TG in patients with acute pancreatitis are (1) marked limitations of dietary intake of fat; (2) the use of low molecular weight heparin or insulin to increase disposal of FFA and TG to tissues; and (3) plasmapheresis, in which TG are filtered from blood or the blood volume is replaced by new blood with lower TG concentrations. However, under circumstances where hepatic TG biosynthesis and secretion is markedly elevated, these approaches have limited value.

An intravenous formulation of diazoxide (Hyperstat®) is known to be used to treat malignant hypertension. The formulation is a sterile solution containing 15 mg/ml of diazoxide at pH 11.5. Administration typically involves the administration of a 300 mg bolus over 5-15 seconds. In some instances, a 150 mg bolus is used, and occasionally the administration is spread over 1 minute. The objective of administration is to rapidly achieve a high free drug concentration in the peripheral arterioles, resulting in extensive vasodilation and rapid reduction in blood pressure. In contrast, intravenous administration of 1.5 mg/kg over 5 minutes has been shown to have little effect on blood pressure (Wang et al. Eur J CardioThoracic Surg, 2003, 24:967-973).

Administration of intravenous K_(ATP) channel agonist, e.g. diazoxide or a salt thereof, in combination with insulin will result in both the reduction of hepatic TG biosynthesis and secretion and increased disposal of circulating TG and FFA to tissues, more effectively lowering circulating concentrations of both than would the use of either drug separately. Additionally, the combination of an intravenous K_(ATP) channel agonist with insulin and reduced dietary intake of fat maximizes reductions in circulating TG and FFA in patients with acute pancreatitis and elevated TG because under these conditions, new chylomicrons are not being synthesized, hepatic TG biosynthesis is suppressed and the disposal to tissue is maximized. As administration of diazoxide partially suppresses insulin secretion and thus raises fasting and post-prandial glucose, insulin supplementation will also inhibit a rise in glucose in patients undergoing intravenous K_(ATP) channel agonist therapy as described herein.

An exemplary intravenous treatment may be as follows. First, a K_(ATP) channel opener, such as diazoxide, may be administered as a bolus at a dose of between about 0.5 to 5 mg/kg over 5 to 10 minutes. Such a bolus administration may result in administration of between about 50 to 200 mg over the administration time. Following the initial bolus, a K_(ATP) channel opener, such as diazoxide, may be administered intravenously at a rate of between about 0.02 to 0.2 mg/kg/hr. During this second K_(ATP) channel opener administration, insulin is administered on a recurring basis at a level and rate sufficient to maintain fasting glucose between about 70 mg/dL and 125 mg/dL and peak post prandial glucose less than 200 mg/dL.

Plasmapheresis may also be used in conjunction with co-administration of diazoxide choline and insulin. Even under circumstances in which plasmapheresis is used, dietary fat intake needs to be restricted to limit the synthesis of new chylomicrons. Additionally, when plasmapheresis is used, hepatic TG biosynthesis and secretion needs to be down-regulated. Therefore the combination of an intravenous K_(ATP) channel agonist with insulin supplementation and plasmapheresis is also a preferred means to treat patients with AP and elevated TG.

Additionally, intravenous administration of a K_(ATP) channel agonist described herein, e.g. diazoxide, has the potential to protect pancreatic and peripancreatic tissues from ischemia and other cytotoxic insult, limiting the potential for complications, infection and organ failure, and thereby also reducing mortality risk.

Nicorandil (2-[(pyridin-3-ylcarbonyl)amino]ethyl nitrate) is a vasodilatory drug combining nitrate functionality with K_(ATP) channel activation. By virtue of its K_(ATP) channel activity, nicorandil has been shown to protect tissue from ischemic injury. Nicorandil may be administered either as an oral product or by intravenous injection. In order to protect tissues in AP patients with elevated TG from ischemic injury and necrosis, it is possible to co-administer nicorandil and diazoxide or a salt thereof, or to develop an intravenous co-formulation of nicorandil and diazoxide or a salt thereof. Nicorandil has not been demonstrated to suppress hepatic TG biosynthesis and secretion to the degree that diazoxide does, but ischemic protection may be improved.

Intravenous administration of a K_(ATP) channel agonist described herein, e.g. diazoxide, is easily achieved during the early stages of hospitalization and may continue while patients are in the intensive care unit. It becomes more challenging once they become ambulatory again. Therefore it is optimal to treat patients with AP and elevated triglycerides with intravenous formulations prior to them becoming ambulatory, and transitioning to oral administration of a K_(ATP) channel agonist described herein, e.g. diazoxide choline, thereafter. In some instances, patients may be treated orally with a K_(ATP) channel agonist described herein, e.g. diazoxide choline, only while they are hospitalized. Alternatively, they may receive a K_(ATP) channel agonist described herein orally on a chronic basis for as long as they have risk of recurrence of AP. Oral administration of a K_(ATP) channel agonist described herein may be achieved with suitable delivery form, including oral suspension, fast-melt tab, immediate release capsule or tablet, controlled release capsule or tablet formulations, etc. Optimally, the patient is treated with a controlled-release formulation which facilitates once per day (QD) dosing and provides for high bioavailability of the K_(ATP) channel agonist.

Kits in the Treatment of Dyslipidemia

Compliance in the treatment of a number of disease conditions requiring treatment with two or more medications, which have not or cannot be co-formulated, is improved if the medications are combined in a kit which includes clear instructions for treatment. Improved compliance results in improved therapeutic outcomes and reduced risk of disease recurrence or relapse. The use of kits to improve compliance in the treatment of chronic asymptomatic diseases is important because the patient is, by definition, unaware or less aware of their disease status. Examples of chronic asymptomatic diseases include hypertension and most lipid abnormalities. Additionally, improved compliance is important to reduce the risk of an uncommon or rare acute effect of the chronic condition. For example, treatments to reduce very high triglycerides are intended to reduce the risk of pancreatitis. Similarly, treatments to reduce elevated LDL are intended to reduce the risk of major cardiovascular adverse events including myocardial infarction and stroke, and to reduce mortality.

In one example a kit is prepared for the treatment of a patient with a lipid disorder characterized by elevated LDL-C and elevated triglycerides which includes a monthly dose of an oral formulation of a K_(ATP) channel agonist, a monthly dose of a statin, and instructions for the administration of each drug. In this kit, the K_(ATP) channel agonist may be a K_(ATP) channel agonist of Formulae I-IV, or a salt thereof. For example, the K_(ATP) channel agonist may be diazoxide choline. In some examples, the diazoxide choline may be in a controlled, delayed, or sustained release formulation. In some examples, the K_(ATP) channel agonist and the statin may be included as separate formulations. Alternatively, the K_(ATP) channel agonist and statin may be co-formulated, e.g., in a single capsule, pill, or other oral delivery form.

In another example a kit is prepared for the treatment of a patient with a lipid disorder characterized by elevated triglycerides which includes a monthly dose of an oral formulation of a K_(ATP) channel agonist, a monthly dose of a fibrate, and instructions for the administration of each drug. In this kit, the K_(ATP) channel agonist may be a K_(ATP) channel agonist of Formulae I-IV, or a salt thereof. For example, the K_(ATP) channel agonist may be diazoxide choline. In some examples, the diazoxide choline may be in a controlled, delayed, or sustained release formulation. In some examples, the fibrate may be fenofibrate or a salt thereof. In some examples, the K_(ATP) channel agonist and the fibrate may be included as separate formulations. Alternatively, the K_(ATP) channel agonist and fibrate may be co-formulated, e.g., in a single capsule, pill, or other oral delivery form.

In yet another example a kit is prepared for the treatment of a patient with a lipid disorder characterized by elevated triglycerides which includes a monthly dose of an oral formulation of a K_(ATP) channel agonist, a monthly dose of an omega-3 fatty acid-based drug, and instructions for the administration of each drug. The kit includes a monthly dose of an oral formulation of a K_(ATP) channel agonist, a monthly dose of a statin, and instructions for the administration of each drug. In this kit, the K_(ATP) channel agonist may be a K_(ATP) channel agonist of Formulae I-IV, or a salt thereof. For example, the K_(ATP) channel agonist may be diazoxide choline. In some examples, the diazoxide choline may be in a controlled, delayed, or sustained release formulation. In some examples, the K_(ATP) channel agonist and the omega-3 fatty acid-based drug may be included as separate formulations. Alternatively, the K_(ATP) channel agonist and omega-3 fatty acid-based drug may be co-formulated, e.g., in a single capsule, pill, or other oral delivery form. The omega-3 fatty acid-based drug may be based on Lovaza®. The omega-3 fatty acid-based drug may be in a controlled, delayed, or sustained release formulation. The omega-3 fatty acid based drug may contain 90% or higher EPA and no DHA.

In yet another example, a kit is prepared for the treatment of a patient with a lipid disorder characterized by reduced HDL-C and elevated triglycerides which includes a monthly dose of an oral formulation of a K_(ATP) channel agonist, a pharmaceutical formulation of niacin or an analog thereof, and instructions for the administration of each drug. In this kit, the K_(ATP) channel agonist may be a K_(ATP) channel agonist of Formulae I-IV, or a salt thereof. For example, the K_(ATP) channel agonist may be diazoxide choline. In some examples, the diazoxide choline may be in a controlled, delayed, or sustained release formulation. In some examples, the K_(ATP) channel agonist and the niacin or analog thereof may be included as separate formulations. Alternatively, the K_(ATP) channel agonist and niacin or analog thereof may be co-formulated, e.g., in a single capsule, pill, or other oral delivery form.

The invention will now be described with reference to the following non-limiting examples.

Examples A. Potassium ATP Channel Activator Containing Formulations 1. Compressed Tablet Formulations of Diazoxide Salt or Derivative

Diazoxide salt or a derivative thereof at about 15-30% by weight is mixed with hydroxypropyl methylcellulose at about 55-80% by weight, ethylcellulose at about 3-10 wt/vol % and magnesium stearate (as lubricant) and talc (as glidant) each at less than 3% by weight. The mixture is used to produce a compressed tablet as described in Reddy et al., AAPS Pharm Sci Tech 4(4):1-9 (2003). The tablet may be coated with a thin film as discussed below for microparticles.

A tablet containing 100 mg of diazoxide salt or a derivative thereof will also contain approximately 400 mg of hydroxypropyl cellulose and 10 mg of ethylcellulose. A tablet containing 50 mg of diazoxide salt or a derivative thereof will also contain approximately 200 mg of hydroxypropyl cellulose and 5 mg of ethylcellulose. A tablet containing 25 mg of diazoxide salt or a derivative thereof will also contain approximately 100 mg of hydroxypropyl cellulose and 2.5 mg of ethylcellulose.

2. Encapsulated Coated Microparticle Formulation of Diazoxide Salt or Derivative

Diazoxide salt or a derivative thereof is encapsulated into microparticles in accordance with well known methods (see, e.g. U.S. Pat. No. 6,022,562). Microparticles of between 100 and 500 microns in diameter containing diazoxide salt or a derivative thereof, alone or in combination with one or more suitable excipient, is formed with the assistance of a granulator and then sieved to separate microparticles having the appropriate size. Microparticles are coated with a thin film by spray drying using commercial instrumentation (e.g. Uniglatt Spray Coating Machine). The thin film comprises ethylcellulose, cellulose acetate, polyvinylpyrrolidone and/or polyacrylamide. The coating solution for the thin film may include a plasticizer which may be castor oil, diethyl phthalate, triethyl citrate and salicylic acid. The coating solution may also include a lubricating agent which may be magnesium stearate, sodium oleate, or polyoxyethylenated sorbitan laurate. The coating solution may further include an excipient such as talc, colloidal silica or of a mixture of the two added at 1.5 to 3% by weight to prevent caking of the film coated particles.

3. Formulations for Controlled Release of Diazoxide or Derivative

3.1. Formulation of a Tableted Form of Diazoxide or a Derivative for Controlled Release

Prior to mixing, both the active ingredient and hydroxypropyl methylcellulose (Dow Methocel K4M P) are passed through an ASTM 80 mesh sieve. A mixture is formed from 1 part diazoxide salt or a derivative thereof to 4 parts hydroxypropyl methylcellulose. After thorough mixing, a sufficient volume of an ethanolic solution of ethylcellulose as a granulating agent is added slowly. The quantity of ethylcellulose per tablet in the final formulation is about 1/10th part. The mass resulting from mixing the granulating agent is sieved through 22/44 mesh. Resulting granules are dried at 40° C. for 12 hours and thereafter kept in a desiccator for 12 hours at room temperature. Once dry the granules retained on 44 mesh are mixed with 15% fines (granules that passed through 44 mesh). Talc and magnesium stearate are added as glidant and lubricant at 2% of weight each. A colorant is also added. The tablets are compressed using a single punch tablet compression machine.

3.2. Formulation of a Compression Tableted Form of Diazoxide or a Derivative Thereof that Provides for Controlled Release.

Diazoxide salt or a derivative thereof at 20-40% weight is mixed with 30% weight hydroxypropyl methylcellulose (Dow Methocel K100LV P) and 20-40% weight impalpable lactose. The mixture is granulated with the addition of water. The granulated mixture is wet milled and then dried 12 hours at 110° C. The dried mixture is dry milled. Following milling, 25% weight ethylcellulose resin is added (Dow Ethocel 10FP or Ethocel 100FP) followed by 0.5% weight magnesium stearate. A colorant may also be added. The tablets are compressed using a single punch tablet compression machine (Dasbach et al., Poster at AAPS Annual Meeting Nov. 10-14 (2002)).

3.3. Formulation of a Compression Coated Tableted Form of Diazoxide or a Derivative Thereof that Provides for Controlled Release.

The core tablet is formulated by mixing either 100 mg of diazoxide salt or a derivative thereof with 10 mg of ethylcellulose (Dow Ethocel 10FP), or by mixing 75 mg of diazoxide or a derivative thereof with 25 mg lactose and 10 mg of ethylcellulose (Dow Ethocel 10FP), or by mixing 50 mg of diazoxide or a derivative thereof with 50 mg of lactose and 10 mg of ethylcellulose (Dow Ethocel 10FP). The core tablets are formed on an automated press with concave tooling. The compression coating consisting of 400 mg of polyethylene oxide (Union Carbide POLYOX WSR Coagulant) is applied and compressed to 3000 psi (Dasbach et al., Poster at AAPS Annual Meeting Oct. 26-30 (2003)).

3.4. Formulation of a Controlled Release Tableted Form of Diazoxide Choline Salt

3.4.1 Controlled Release Formulations

Controlled release tableted formulations of diazoxide choline salt were developed and investigated with respect to a variety of properties known by those of skill in the pharmaceutical art relating to the manufacture of tableted formulations including, for example, ease and consistency of manufacture, appearance (e.g., sheen, compressibility, microscopic appearance), and dissolution properties (e.g., rate, order and extent of dissolution). Tablets were produced individually on a press, where the final blend of diazoxide choline salt and excipient was weighed out to the desired total tablet weight prior to compression. As shown in Table 2, Formulations A-H, J, and L contained 50.0 mg diazoxide as the choline salt (i.e., 72.5 mg total diazoxide choline salt present), Formulations I and K contained 200.0 mg diazoxide as the choline salt (i.e., 290.0 mg total diazoxide choline salt present) and Formulation U contained 145 mg diazoxide as the choline salt. The manufacture of formulation L is exemplary of the manufacturing methods available to the skilled artisan. For formulation L, diazoxide choline salt, talc, and approximately half of the colloidal silicon dioxide (Cab-o-sil) were mixed in a KG-5 mixer bowl with an impeller speed of about 300 rpm and a chopper speed of about 3000 rpm for about 4 min. The mixture was passed through a co-mil equipped with a 024R screen, square-edged paddle, and 0.175″ spacer. To this milled mixture in a 8-qt V-shell blender was added Emcompress (dibasic calcium phosphate) through a #20 mesh hand screen with blending for about 10 min. To this mixture was added PEO N750 and PEO 303, which had been passed through a #20 mesh hand screen, with blending for about 10 min. To this mixture was added Pruv and the remainder of the Cab-o-sil, having been passed through a #20 mesh hand screen, with blending for about 5 min. The mixture was subjected to pressing (Manesty Beta Press) using 0.2220″×0.5720″ caplet shaped tooling (Set #21.)

TABLE 2 Exemplary Formulations for Diazoxide Choline salt. Formulation INGREDIENT Amount per tablet (mg) A. Diazoxide Choline 72.50 Kollidon SR 100.0 PEO N303 25.00 Emcompress 50.50 Pruv 2.00 TOTAL WEIGHT 250.0 B. Diazoxide Choline 72.50 Methocel K100M 237.3 Emcompress 35.00 Magnesium Stearate 3.50 Cab-o-sil 1.75 TOTAL WEIGHT 350.0 C. Diazoxide Choline 72.50 Kollidon SR 105.0 PEO N303 35.0 Emcompress 134.1 Pruv 3.50 TOTAL WEIGHT 350.1 D. Diazoxide Choline 72.50 Methocel K100M 175.1 Emcompress 97.30 Magnesium Stearate 3.50 Cab-o-sil 1.75 TOTAL WEIGHT 350.2 E. Diazoxide Choline 72.50 PEO N750 NF 105.0 PEO N303 NF 52.50 Emcompress 116.6 Pruv 3.50 TOTAL WEIGHT 350.1 F. Diazoxide Choline 72.50 Kollidon SR 105.0 PEO N303 7.00 Emcompress 162.1 Pruv 3.50 TOTAL WEIGHT 350.1 G. Diazoxide Choline 72.50 Methocel K100M 175.1 Emcompress 105.1 Magnesium Stearate 3.50 Cab-o-sil 1.75 TOTAL WEIGHT 358.0 H. Diazoxide Choline 72.50 PEO N750 NF 105.0 PEO N303 NF 35.01 Emcompress 134.1 Pruv 3.50 TOTAL WEIGHT 350.1 I. Diazoxide Choline 290.0 Methocel K100M 240.0 Emcompress 258.0 Magnesium Stearate 8.00 Cab-o-sil 4.00 TOTAL WEIGHT 800.0 J. Diazoxide Choline 72.50 PEO N750 NF 105.0 PEO N303 NF 52.50 Emcompress 116.6 Pruv 3.50 Talc 3.50 Cab-o-sil 1.75 TOTAL WEIGHT 355.4 K. Diazoxide Choline 290.0 PEO N750 NF 249.0 PEO N303 NF 124.5 Emcompress 145.3 Pruv 8.30 Talc 8.30 Cab-o-sil 4.15 TOTAL WEIGHT 829.6 K′. Diazoxide Choline 290.0 PEO N750 NF 249.0 PEO N303 NF 124.5 Emcompress 137.0 Pruv 8.30 Talc 16.6 Cab-o-sil 4.15 TOTAL WEIGHT 830.0 L. Diazoxide Choline 72.51 PEO N750 NF 105.1 PEO N303 NF 52.54 Emcompress 111.4 Pruv 3.50 Talc 3.50 Cab-o-sil 1.75 TOTAL WEIGHT 350.3 U. Diazoxide Choline 145 Pregelatinized starch 177 PEO N750 NF 20 PEO N303 NF 10 Microcrystalline 230 cellulose NF Colloidal silicone 6 dioxide NF Magnesium stearate 12 NF TOTAL WEIGHT 600

It is noted that the composition of formula U in Table 2 is for the core tablet. The core tablet U was coated with a 2% opadry clear coat followed by a 3% Surelease coating.

Microscopic observation of the tablets having Formulation A revealed a grainy texture of the diazoxide choline salt, and at the 29% loading of diazoxide choline salt in Formulation A the blend had poor flow characteristics. Accordingly, the loading of diazoxide choline was reduced in Formulation B. A caplet shaped tooling, approximately 6 mm×15 mm was found to result in acceptable tablet appearance, sheen, and ease of compressibility. However, Formulation B also exhibited poor blend flow.

Subsequent formulations (e.g., Formulations C-L) incorporated a milling step of the diazoxide choline salt prior to incorporation into the tablet blend. A milling study was conducted using a test mill equipped with different screen sizes to evaluate and determine a suitable milling process. Particle size was determined by visual comparison with 40 m reference beads. As shown in Table 3.1, use of the 024R screen resulted in the best recovery of API (i.e., “active pharmaceutical ingredient”=diazoxide as the diazoxide choline salt) providing the broadest range of particle sizes. Accordingly, material milled through a 024R screen was selected for subsequent formulation.

TABLE 3.1 Milling study for diazoxide choline salt prior to formulation API loaded/API recovered Particle size Screen/Paddle/Spacer/Speed (g) (μm) 018R/square/0.200″/80% 50.0/36.5 ~100-200 024R/square/0.200″/80% 50.0/42.0 ~100-250 039R/square/0.200″/80% 50.0/42.0 ~150-250

Further optimization of formulations containing the diazoxide choline salt were carried out. Previous PEO formulations showed signs of sticking to the punch surface, possibly due to the interaction of PEO and diazoxide choline. Five additional formulations were prepared using polymers such as polyethylene oxide and cellulose in an attempt to discover non-sticking formulations.

Formulations M-P and R-S were prepared at a 500 g scale with the compositions shown in Table 3.2. Formulation Q was prepared as a small batch of five hand-compressed tablets. Formulation T was prepared at the 2 kg scale. In formulation M, the amount of PEO was reduced from 45% to 25%. In formulation N, PEO was replaced with 25% hydroxypropylmethyl cellulose (HPMC) and microcrystalline cellulose was added. Formulation 0 was similar to N, but used a different type of HPMC and dropped the starch. In formulation P, less HPMC and more microcrystalline starch were used than in N, and the starch was absent. Formulation Q was identical to P except that a different type of HPMC was used. Formulations R, S, and T included higher loadings of diazoxide choline (290 mg) and reduced tablet size (400, 700, and 830 mg for R, S, and T, respectively).

Formulations M-T were according to the following general procedure. Diazoxide tablet blend procedure. Diazoxide choline, colloidal silicon dioxide and half the talc were mixed in a v-shell blender for three minutes. The mixture was then co-milled for particle size reduction and to improve flow. The milled diazoxide choline mixture was blended with the remaining talc and either microcrystalline cellulose or starch in a v-shell blender for three minutes. The resulting mixture was subjected to roller compaction with a #12 mesh in line mill to provide granules. The granules were sieved through #16 and #40 mesh sieves. Material retained by the #16 mesh and passing through the #40 mesh were passed through the compacetor again. Properly sized granules were roller compacted with half of the magnesium stearate and blended for three minutes in a v-shell blender. PEO and dibasic calcium phosphate were blended with the resulting mixture for three minutes in a v-shell blender. The remaining magnesium stearate was blended into the mixture for two minutes using a v-shell blender.

All the tablets produced had a nice appearance with good clean edges and most likely could easily withstand the coating process. No signs of sticking or picking were present. The roller compaction method worked well for these formulations. Tablets prepared from formulations N, O, and P reached a maximum hardness of about 8 kp, while tablets of formulation N reached a maximum hardness of about 2.8 to 3.0 kp; the latter may have been due to obtaining a maximum of 3 tons of compression during manufacture. Formulations S and T showed signs of lamination, but that may be overcome by, e.g., precompression, use of a different roller compaction screen, and increasing the amount of fines in final blend.

TABLE 3.2 Additional Diazoxide Choline Salt Formulations FORMULATION (wt %) INGREDIENT M N O P Q Diazoxide Choline 34.94 34.90 34.90 34.90 34.90 Polyethylene Oxide, 17.00 — — — — N750 Polyethylene Oxide, 8.00 — — — — 303 Methocel A15 — — 25.00 15.00 Methocel K100M — 25.00 15.00 Avicel PH101 — 10.00 9.50 19.50 19.50 Emcompress — 24.60 24.60 24.60 24.60 Talc, USP 2.00 2.00 2.00 2.00 2.00 Starch 1500 34.06 16.78 — — — Magnesium Stearate 3.00 3.00 3.00 3.00 3.00 Cab-o-sil M5P 1.00 0.50 1.00 1.00 1.00 Total 100.0 100.0 100.0 100.0 100.0 FORMULATION (wt %) INGREDIENT R S T Diazoxide Choline 72.5 41.43 34.90 Polyethylene Oxide, 2.00 10.00 17.00 N750 Polyethylene Oxide, 1.00 5.00 8.00 303 Methocel A15 — — — Methocel K100M — — — Avicel PH101 — — — Emcompress — — — Talc, USP 2.00 2.00 2.00 Starch 1500 18.5 37.57 34.06 Magnesium Stearate 3.00 3.00 3.00 Cab-o-sil M5P 1.00 0.50 1.00 Total 100.0 100.0 100.0

3.4.2. Dissolution Studies

Dissolution of tablets with formulation as set forth in Table 4 was investigated. One or more tablets (e.g., 1 or 2) of the indicated tableted formulation were placed into a volume of buffer (e.g., 900 mL) with known buffer salt concentration (e.g., 0.05 M potassium phosphate, pH 8.6; 0.05 M potassium phosphate, pH 7.5), with or without surfactant (e.g., 0.05% CTAB), at a known temperature (e.g., 37° C.). Stirring conditions employed paddles at, for example, 50 RPM. Aliquots (e.g, 10 mL) removed as a function of time were filtered (e.g., 0.45 m GMF Filter) prior to analysis.

TABLE 4 Dissolution study for formulations of diazoxide choline salt (entries are % dissolved diazoxide) Time (hr) Entry Protocol 0 1 2 3 6 9 12 18 24 C a 0.0 2.9 4.5 5.7 9.4 13.0 16.6 D a 0.0 4.5 7.8 11.0 19.1 25.6 31.3 F b 29 36 44 54 G b 30 39 50 67 H b 50 75 91 101 I b 25 37 48 57 J b 34 53 76 104 K b 25 43 64 94 24 41 62 92 U b 0 6 32 77 101 M b 18 34 57 90 N b 56 O b 23 42 61 71 P b 22 37 51 65 R b 102 104 105 106 Q b 27 45 62 74 S b 30 56 85 106 T b 24 44 73 99 Column 1: Entry from Table 2. Column 2: Protocol a) 0.05M potassium phosphate, pH 8.6, 37° C.; Protocol b) 0.05M potassium phosphate, pH 7.5, 0.05% CTAB, 37° C.

Subsequently, 200 mg tablets of formulations R, S and T were coated with a 1-2% clearcoat undercoat and with 1-7% ethycel (Surelease) overcoat. This combination of undercoat and Surelease overcoat delayed release of drug from the formulation by approximately 1 hour followed by linear release through 24 hours. Adjustment of the Surelease concentration from 7% to 3% results in increases in the rate of release without impacting the linearity of release. For example formula R with a polymeric undercoat and 7% surelease coat results in 61% release by 24 hours using protocol b (See Table 4.1). Similarly, a 600 mg tablet of formulation U (core tablet) was coated with a 2% opadry clear coat, and then with a 3% Surelease coating.

TABLE 4.1 Dissolution of Coated Tablets Formulations at 7%, 5%, and 3% Overcoating Time (h) R 7% S 5% T 3% 0 0 0 0 1 0 0 1 3 2 4 5 6 8 13 15 12 25 36 39 24 61 73 81

The dissolution profile (i.e., % dissolved with time) of Proglycem® capsules (100 mg) and controlled-release tablet formulations of diazoxide as provided herein is shown in Table 5. The 50 mg tablet entry of Table 5 refers to Formulation J (Table 2) wherein talc and cab-o-sil was present at 2% and 1%, respectively. The 200 mg tablet entry of Table 5 refers to Formulation K (Table 2) wherein talc and cab-o-sil are present at 2% and 1%, respectively.

TABLE 5 Dissolution profile of 100 mg Proglycem ® (capsule) and diazoxide choline salt (tablet) 200 mg tablet 50 mg tablet 100 mg capsule diazoxide choline diazoxide choline Time (hr) Proglycem ® salt salt 0 0 0 0 3 79 22 25 6 85 36 39 12 92 54 70 24 98 88 99

As evidenced by Table 5, the 100-mg Proglycem® capsule provides for faster dissolution of diazoxide relative to tablets described herein. Approximately 79% diazoxide component of Proglycem® is recovered in dissolution buffer after 3-hr. In contrast, the 50 and 200-mg tablets described in Table 5 dissolved at levels of 25% and 22%, respectively, at 3-hr. At 12-hr, 100-mg Proglycem® capsule dissolved at 92%, whereas the 50 and 200-mg tablets dissolved at 70% and 54%, respectively. Approximately total dissolution is observed with 100-mg Proglycem® capsule and 50-mg tablet at 24 hrs.

3.4.3. Excipient Compatibility Studies

Studies to determine the compatibility of diazoxide choline salt for various excipients are summarized in Table 6. Each mixture of excipient (100 mg) and diazoxide choline salt (100 mg) was made in acetonitrile to 10 mL Samples were assayed immediately (i.e., “Initial” column of Table 6) and at one-month storage under conditions a) 40° C./75% RH (relative humidity), and b) 50° C.

TABLE 6 Excipient compatibility study for diazoxide choline salt. 1 Month 1 Month Initial % 40° C./75% RH % 50° C. % Excipient (w/w) (w/w) (w/w) Hydroxyrpropylmethyl cellulose (HMPC) 98.8 101.4 96.6 Hydroxypropylcellulose (HPC) 103.1 97.4 100.8 Ethylcellulose (EC) 90.6 102.0 99.7 99.5* Methylcellulose (MC) 102.3 100.3 99.1 Carboxymethyl cellulose Na (CMC Na) 96.8 101.6 99.5 Starch 1500 103.3 90.6 95.2 Kollidon SR 81.2 99.8 100.9 100.2* Polyethyleneoxide N3 03 (PEO) 81.0 99.2 99.0 99.1* Dibasic Calcium Phosphate 98.4 102.9 102.2 Sodium Stearyl Fumarate 98.9 101.1 103.7 Magnesium Stearate 100.1 101.4 99.3 Colloidal Silicon Dioxide (Cab-o-sil) 99.0 99.4 104.5 Microcrystalline Cellulose 96.8 98.6 107.0 Lactose Monohydrate 94.5 99.2 103.0 Mannitol 111.7 98.1 81.3 Diazoxide Control N/A 99.0 99.4 *Re-assayed

Initial low results for ethylcellulose, Kollidon SR, and polyethyleneoxide were investigated by re-preparing samples of diazoxide choline salt and excipient. As shown in Table 6, initial sample recovery (i.e., column “Recovery”) of the re-prepared samples indicates method accuracy.

No reportable impurities were detected in any sample, and no degradation was observed in the samples stored for one-month with the exception of mannitol wherein 81.3% of diazoxide was recovered after 1-month at 50° C.

3.4.3. Stability Studies for Diazoxide Choline Controlled Release Tablets

Studies to determine the stability of formulations of diazoxide choline salt as the controlled release tablet were conducted on sample formulations as described in Table 2, results of which are shown in Table 7. In these studies, storage conditions were the following: a) 25° C./60% RH, and b) 40° C./75% RH. In Table 7, the term “Appearance” refers to the physical appearance of the tablets of the study. The term “Assay (%)” refers to the percentage of diazoxide choline salt assayed with respect to nominal (i.e., 50 mg or 200 mg) content of the sample The term “Dissolution (%)” refers to the amount, expressed as a percentage, of diazoxide choline salt assayed by method described herein.

TABLE 7 Stability Study for Formulations of Diazoxide Choline Salt. Time point Test Initial 1 month 2 months 50 mg tablet, Formulation J, 25° C./60% RH, Appearance White tablets White tablets White tablets Assay (%) 95.8 102.9 99.3 Dissolution (%):  3 hr 24 19 15  6 hr 41 30 27 12 hr 62 48 52 24 hr 92 90 94 50 mg tablet, Formulation J, 40° C./75% RH, Appearance White tablets White tablets White tablets Assay (%) 95.8 103.2 99.4 Dissolution (%):  3 hr 24 10 10  6 hr 41 22 21 12 hr 62 50 46 24 hr 92 90 84 200 mg tablet, Formulation K, 25° C./60% RH Appearance White tablets White tablets White tablets Assay (%) 95.9 100.9 100.3 Dissolution (%):  3 hr 34 15 13  6 hr 53 31 30 12 hr 76 69 68 24 hr 104 101 96 200 mg tablet, Formulation K, 40° C./75% RH Appearance White tablets White tablets White tablets Assay (%) 95.9 99.6 100.0 Dissolution (%):  3 hr 34 10 10  6 hr 53 26 28 12 hr 76 60 61 24 hr 104 90 90

3.5. A Controlled Release Dosage Form of Diazoxide or a Derivative Thereof Using an Osmotically Controlled Release System.

Diazoxide salt or a derivative thereof is formulated as an osmotically regulated release system. In general, two components, and an expandable hydrogel that drives release of the active drug is assembled with diazoxide salt or a derivative thereof into a semipermeable bilaminate shell. Upon assembly a hole is drilled in the shell to facilitate release of active upon hydration of the hydrogel.

A dosage form adapted, designed and shaped as an osmotic delivery system is manufactured as follows: first, a diazoxide salt or a derivative thereof composition is provided by blending together into a homogeneous blend of polyethylene oxide, diazoxide salt or a derivative thereof and hydroxypropyl methylcellulose. Then, a volume of denatured anhydrous ethanol weighing 70% of the dry mass is added slowly with continuous mixing over 5 minutes. The freshly prepared wet granulation is screened through a 20 mesh screen through a 20 mesh screen, dried at room temperature for 16 hours, and again screened through a 20 mesh screen. Finally, the screened granulation is mixed with 0.5% weight of magnesium stearate for 5 minutes.

A hydrogel composition is prepared as follows: first, 69% weight of polyethylene oxide weight, 25% weight of sodium chloride and 1% weight ferric oxide are separately screened through a 40 mesh screen. Then, all the screened ingredients are mixed with 5% weight of hydroxypropyl methylcellulose to produce a homogeneous blend. Next, a volume of denatured anhydrous alcohol equal to 50% of the dry mass is added slowly to the blend with continuous mixing for 5 minutes. The freshly prepared wet granulation is passed through a 20 mesh screen, allowed to dry at room temperature for 16 hours, and again passed through a 20 mesh screen. The screened granulation is mixed with 0.5% weight of magnesium stearate for 5 minutes (see U.S. Pat. No. 6,361,795 by Kuczynski, et al.).

The diazoxide salt composition, or a derivative thereof, and the hydrogel composition, are compressed into bilaminate tablets. First the diazoxide salt or a derivative thereof composition is added and tamped. The hydrogel composition is then added and the laminae are pressed under a pressure head of 2 tons into a contacting laminated arrangement.

The bilaminate arrangements are coated with a semipermeable wall (i.e. thin film). The wall forming composition comprises 93% cellulose acetate having a 39.8% acetyl content, and 7% polyethylene glycol. The wall forming composition is sprayed onto and around the bilaminate.

Finally, an exit passageway can be drilled through the semipermeable wall to connect the diazoxide salt or a derivative thereof drug lamina with the exterior of the dosage system. Residual solvent is removed by drying at 50° C. and 50% humidity. The osmotic systems are dried at 50° C. to remove excess moisture (see U.S. Pat. No. 6,361,795 by Kuczynski, et al.).

4. Preparation of Diazoxide Salts

4.1. Preparation of the Sodium Salt

The sodium salt of diazoxide was prepared by dissolving 300 mg of diazoxide in approximately 45 mL methyl ethyl ketone (MEK). The diazoxide/MEK solution was heated at 75° C. on an orbital shaker to ensure dissolution. To the solution was added 1.3 mL of 1M NaOH (1 molar equivalent). The combined solutions were heated at 75° C. for approximately 30 minutes and allowed to cool to room temperature. The mixture was concentrated under reduced pressure, and dried in vacuo at 55° C. and 30 in·Hg. Elemental analysis: Calculated, 38.03% C, 2.39% H, 11.09% N and 9.1% Na; Found, 38.40% C, 2.25% H, 10.83% N and 7.4% Na.

A sodium salt of diazoxide was also prepared by dissolving 300 mg of diazoxide in approximately 45 mL acetonitrile. The diazoxide/acetonitrile solution was heated at 75° C. on an orbital shaker to ensure dissolution. To the solution was added 1.3 mL of 1M NaOH (approximately 1 molar equivalent). The combined solutions were heated at 75° C. for approximately 30 minutes and allowed to cool to room temperature. The mixture was concentrated under reduced pressure, and dried in vacuo at 55° C. and 30 in·Hg.

4.2. Preparation of the Potassium Salt

The potassium salt of diazoxide was prepared by dissolving 300 mg of diazoxide in approximately 45 mL methyl ethyl ketone (MEK). The diazoxide/MEK solution was heated at 75° C. on an orbital shaker to ensure dissolution. To the solution was added approximately 1.3 mL of 1 M KOH (1 molar equivalent) and the solution was returned to the orbital shaker, heated at 75° C. for approximately 30 minutes and allowed to cool to room temperature. The solvent was removed under reduced pressure and the solid was dried in vacuo at 55° C. and 30 in·Hg. Elemental analysis: Calculated, 33.59% C, 2.81% H, 9.77% N and 13.63% K; Found, 34.71% C, 2.62% H, 9.60% N and 10.60% K.

The potassium salt of diazoxide was also prepared by dissolving 300 mg of diazoxide in approximately 45 mL tetrahydrofuran (THF). The diazoxide/THF solution was heated at 75° C. on an orbital shaker to ensure dissolution. To the solution was added approximately 1.3 mL of 1 M KOH (1 molar equivalent) and the resulting solution was returned to the orbital shaker, heated at 75° C. for approximately 30 minutes and allowed to cool to room temperature. THF was removed under reduced pressure and the solid was dried in vacuo at 55° C. and 30 in·Hg.

4.3. Preparation of Choline Salt

4.3.1. Preparation of the Choline Salt: Proof of Concept

The choline salt of diazoxide was prepared by dissolving 300 mg of diazoxide in approximately 45 mL methyl ethyl ketone (MEK). The diazoxide/MEK solution was heated at 75° C. on an orbital shaker to ensure dissolution. To the solution was added approximately 315 mg of 50 wt. % of choline hydroxide (1 molar equivalent) and the solution was returned to the orbital shaker and stirred at 75° C. for approximately 30 minutes. The solvent was removed under reduced pressure, and the solid was dried in vacuo at 55° C. and 30 in·Hg. Elemental analysis: Calculated, 46.77% C, 5.86% H, and 12.00% N; Found, 46.25% C, 6.04% H, and 12.59% N.

4.3.2. Process of Preparation of the Choline Salt

An investigation of the minimum solvent volumes required to optimize the formation of the diazoxide choline salt was performed in MeCN, THF, MEK, and 2-MeTHF on small scale in the presence and absence of MTBE. Analyses by 1H NMR and XRPD of all reactions were consistent with the assigned structure and Form B of diazoxide choline salt.

4.3.2.1. Solvent Efficiency Study with Single Solvent System.

Investigation of single solvent system synthesis of diazoxide choline salt using solvents THF, MEK, and 2-MeTHF were conducted, results of which are shown in Table 8. These results suggest that precipitation can be achieved with a single solvent system in THF or 2-MeTHF. The best results, (i.e., Table 8 entries 14-17) obtained with 2-MeTHF. However, it was observed that in 2-MeTHF at 3-volumes and above, complete dissolution was not achieved after addition of choline hydroxide.

TABLE 8 Solvent Efficiency of a Single Solvent System Entry Solvent Solvent volume Yield (%) Polymorphic Form 1 THF 1.0  7 B 2 THF 2.0 n/a n/a 3 THF 3.0 20 B 4 THF 4.0 28 B 5 THF 5.0 32 B 6 THF 6.0 42 B 7 THF 7.0 53 B 8 THF 8.0 50 B 9 MEK 4.0 n/a n/a 10 2-MeTHF 1.0 n/a n/a 11 2-MeTHF 2.0 48 B 12 2-MeTHF 3.0 41 B 13 2-MeTHF 4.0 48 B 14 2-MeTHF 5.0 71 B 15 2-MeTHF 6.0 71 B 16 2-MeTHF 7.0 72 B 17 2-MeTHF 8.0 67 B n/a: Samples were either seen to form a gum, or no precipitate was observed.

4.3.2.2. Solvent Efficiency Study with Binary Solvent Systems.

The addition of a second solvent at 1-20 volumes (i.e., 1-8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20) to the solvent volumes of Table 8 was envisaged to optimize diazoxide salt production yields. Results of investigation of methods to enhance precipitation and increase yield employing binary solvent system synthesis of diazoxide choline salt using solvents THF, MeCN, MEK, and 2-MeTHF and co-solvent MTBE are shown in Table 9. Optimum conditions for the preparation of the diazoxide choline salt obtained with 1-3 volumes of THF (see Table 8, entries 1-3), with 3 volumes being the ratio of choice to eliminate excessive drag during stirring of slurry in large scale production.

TABLE 9 Solvent Efficiency of a Binary Solvent System Solvent Co-Solvent Polymorphic Entry Solvent Volume (12 Vol) Yield (%) Form 1 THF 1.0 MTBE 96 B 2 THF 2.0 MTBE 96 B 3 THF 3.0 MTBE 91 B 4 THF 4.0 MTBE 90 B 5 THF 5.0 MTBE 84 B 6 THF 6.0 MTBE 90 B 7 THF 7.0 MTBE 94 B 8 THF 8.0 MTBE 94 B 9 MeCN 1.0 MTBE 92 B 10 MeCN 2.0 MTBE 90 B 11 MeCN 3.0 MTBE 87 B 12 MeCN 4.0 MTBE 79 B 13 MeCN 5.0 MTBE 79 B 14 MeCN 6.0 MTBE 75 B 15 MeCN 7.0 MTBE 70 B 16 MeCN 8.0 MTBE 74 B 17 MEK 4.0 MTBE 83 B 18 2-MeTHF 1.0 MTBE 97 B 19 2-MeTHF 2.0 MTBE 94 B 20 2-MeTHF 3.0 MTBE 94 B 21 2-MeTHF 4.0 MTBE 95 B 22 2-MeTHF 5.0 MTBE 97 B 23 2-MeTHF 6.0 MTBE 94 B 24 2-MeTHF 7.0 MTBE 97 B 25 2-MeTHF 8.0 MTBE >99 B

4.3.2.3. Co-Solvent Efficiency and Optimization with MTBE.

Results of optimization of co-solvent (MTBE) volume in the presence of 3 volumes of solvent are shown in Table 10.

TABLE 10 Co-Solvent Efficiency and Optimization with MTBE Entry Solvent MTBE Volume Yield (%) Form 1 MeCN 8.0 76 B 2 MeCN 14.0 85 B 3 THF 6.0 93 B 4 THF 8.0 93 B 5 THF 14.0 98 B

Optimized conditions for the production of the choline salt of diazoxide in a binary solvent system obtained with 3:14 (i.e., 1:4.7) THF/MTBE (v/v).

4.3.2.4. Optimization of Cooling Profile.

Optimization for the controlled precipitation of diazoxide choline salt was conducted by varying the cooling temperatures and hold times. Reactions were carried out in MeCN, THF, and 2-MeTHF (3 vol) with MTBE (14 vol). The results are presented in Table 11. These results suggest that optimum crystal growth was achieved with cooling to 0-5° C. for eight hours in THF and 2-MeTHF (Entries 5-6) when compared to those in MeCN (Entries 1,2, and 4). These results however did not indicate enhanced precipitation when compared to previous studies carried out in THF and 2-MeTHF when the slurries were allowed to stir for two hours. In addition no notable improvements were observed for the additional cooling to −15° C. or with the filtration at ambient temperature.

TABLE 11 Cooling Profile Optimization for Controlled Precipitation Cooling Temp. Hold Time Entry Solvent (° C.) (hr) Yield (%) Form 1 MeCN 0-5 2 89 B 2 MeCN −10 → −15 2 85 B 3 THF RT 2 80 B 4 MeCN RT 2 85 B 5 THF 0-5 8 94 B 6 2-MeTHF 0-5 8 95 B

4.3.2.5. Optimization of Rate of Co-solvent addition.

The effect of rate and hold time of co-solvent addition on the precipitation of diazoxide choline salt was examined. The reactions were carried out on 500 mg scale in three volumes of primary solvent and 14 volumes of MTBE (THF and 2-MeTHF) or 1:3 volume ratio (MeCN). Addition of the co-solvent was made dropwise in one portion for Entry 1, and for Entries 2 and 3 the co-solvent was added at a rate of 1 mL/20 minutes for a total hold time of 140 minutes for the addition of 7 mL. The results, presented in Table 12, indicate that optimum precipitation of diazoxide choline salt was achieved by the addition of MTBE with 20 minute hold times in between additions in THF or 2-MeTHF (entries 2-3).

TABLE 12 Optimization of Rate of Co-solvent addition Hold Time Addition Rate Entry Solvent (min) (mL/min) Yield (%) Form 1 MeCN 60 n/a 87 B 2 THF 140 1.0/20 96 B 3 2-MeTHF 140 1.0/20 96 B

4.3.2.6. Thermal Stability Study.

The stability of the isolated diazoxide choline salt was investigated to determine optimal drying conditions to minimize residual solvents without degradation. Samples of the diazoxide choline salt prepared in THF and MTBE (Entries 1-4) were dried under vacuum at 40° C. for various durations on small scale (100 mg). The study was then carried out on 5 g scale (Entry 5) at 30° C. for eight hours under vacuum to show reproducibility on large scale. The results are presented in Table 13. Analyses of the samples of Table 13 by ¹H NMR, XRPD, and HPLC indicated that elevated temperatures and prolonged drying times did not degrade the compound or affect the form of the compound.

TABLE 13 Thermal Stability Study OVI Drying Temp (THF/MTBE) Polymorphic Entry (C.) Time (h) (ppm) Form 1 RT 12 376/248 B 2 40 20 317/160 B 3 40 28 276/127 B 4 40 42 268/128 B 5 30 8 241/509 B

4.3.2.6. Demonstration of 50-g Scale synthesis of Diazoxide Choline Salt in THF.

The preparation of the diazoxide choline salt was carried out in a binary-solvent system of THF and MTBE with a ratio of 1:4.7 (solvent/co-solvent) volumes and cooling to 0-5° C. for two hours with stirring. A demonstration run for the large scale production utilizing the modified procedure was carried out on 50 g-scale. Diazoxide (50 g) as a hot (62° C.) suspension in THF (140 mL) was treated with choline hydroxide (45% solution in MeOH, 1.0 equiv) added (2 mL/min) over 30 minutes. The resulting solution was stirred for 30 minutes, followed by cooling to 52° C. for the addition of MTBE (14 vol) over 45 minutes. On addition of two volumes of co-solvent precipitation was observed. The resultant slurry was then allowed to cool naturally to ambient temperature followed by further cooling to 0-5° C. with an ice/water bath and an additional 2 hr stirring. The precipitate was isolated by vacuum filtration, and the filter cake rinsed with ice cold MTBE (˜50 mL) and dried under vacuum at room temperature for 12 hours. OVI analysis indicated that THF levels were above the recommended ICH guidelines (799 ppm) and the material was returned to the oven at 30° C. for eight hours to give diazoxide choline salt [70.87 g, 97% yield, 97% purity AUC] as a white crystalline solid. Analyses by 1H NMR, XRPD, and HPLC were consistent with the assigned structure of Form B while maintaining a high purity with good yield.

4.3.2.7. Demonstration of 50-g Scale Synthesis of Diazoxide Choline Salt in 2-MeTHF.

A 50-g level synthesis was duplicated in 2-MeTHF/MTBE as an alternative to THF/MTBE for the large scale production of the diazoxide choline salt described above. No issues or concerns arose during the reaction other than complete dissolution was not achieved after addition of the choline hydroxide. A white precipitate formed after addition of the co-solvent, which was isolated via vacuum filtration and dried under vacuum at 30° C. for eight hours to give diazoxide choline salt [71.51 g, 97% yield] as a white crystalline solid. Analyses by 1H, XRPD, and HPLC were consistent with the assigned structure and Form B. OVI analysis showed 2-MeTHF and MTBE levels of 125 and 191 ppm, respectively, which were below the ICH guidelines.

4.3.2.8. 250-g Scale synthesis of Diazoxide Choline Salt in THF.

Larger scale preparation of the diazoxide choline salt was carried out in a binary-solvent system of THF and MTBE with a ratio of 1:4.7 (v/v) volumes. Diazoxide as a hot (62° C.) suspension in THF (745 mL) was treated with choline hydroxide as a 45% solution in MeOH (1.0 equiv) added over 30 minutes. The resulting solution was stirred for 30 minutes, followed by cooling to 52° C. for the addition of MTBE (14 vol) over 45 minutes. On addition of two volumes of co-solvent, precipitation was observed. The resultant slurry was then allowed to cool naturally to ambient temperature followed by cooling to 0-5° C. with an ice/water bath. The precipitate was isolated by vacuum filtration, and the filter cake rinsed with ice cold MTBE (approximately 250 mL) and dried under vacuum at 30° C. for 38 hours to give diazoxide choline salt (350.28 g, 97.7% yield) as a white crystalline solid. Analyses by 1H NMR, XRPD, and HPLC were consistent with the assigned structure of Form B while maintaining a high purity with good yield.

4.3.2.9. 2-kg Scale Synthesis of Diazoxide Choline Salt in THF.

A 12-L reaction flash was charged with 2.0 kg diazoxide and 5.0-L THF with stirring and heating to 55° C. Choline hydroxide (45% solution in methanol, 2.32 L) was added dropwise to this reaction mixture over about 2.5 hr with stirring. The temperature was maintained at 60±5° C. After addition of choline hydroxide, stirring was continued for about 30 min. The reaction mixture was clarified by in-line 10 micron filtration upon transfer to a 22-L reaction flask pre-charged with 2-L pre-filtered THF, into which was added 10-L pre-filtered MTBE dropwise. This reaction mixture was transferred to another flask which was then charged with an additional 30-L pre-filtered MTBE dropwise, with adjustment of temperature to <5° C. and stirring for about 2 hr. Diazoxide choline salt was recovered by vacuum filtration to afford 2.724 kg (94%) diazoxide choline salt (99.8%, HPLC purity), confirmed by 1H NMR, IR, and UV/Visible analysis.

4.4. Preparation of the Hexamethyl Hexamethylene Diammonium Hydroxide Salt

The hexamethyl hexamethylene ammonium salt of diazoxide was prepared by dissolving 50 mg of diazoxide in approximately 7.5 mL methyl ethyl ketone (MEK). The diazoxide/MEK solution was heated at 75° C. on an orbital shaker to ensure dissolution. To the solution was added approximately 2.17 mL of 0.1M hexamethyl hexamethylene ammonium hydroxide solution (1 molar equivalent) and the solution was stirred at 75° C. for an additional 10 minutes and then cooled to room temperature at the rate of 30° C./h. The solvent was removed under reduced pressure, and the solid was dried in vacuo at 55° C. and 30 in·Hg.

4.5. Failure to Obtain Salts of Diazoxide and derivatives

4.5.1. Failure to Obtain Salts from Alkali Metal Hydroxides

U.S. Pat. No. 2,986,573 (“the '573 patent”) describes the synthesis of diazoxide metal salts in aqueous or non-aqueous solutions in the presence of an alkali metal alkoxide. According to the '573 patent, diazoxide can be dissolved in an alkali metal solution, and the salt obtained upon evaporation. Also described is a method for forming salts from non-aqueous media wherein diazoxide and sodium methoxide are dissolved in anhydrous methanol and the solvent is evaporated to obtain the sodium salt of diazoxide as a white solid.

Attempts were made to prepare a diazoxide salt from alkali metal hydroxides by the method described in the '573 patent. Salt preparation was carried out in aqueous media by dissolving diazoxide in a basic solution 1M NaOH, followed by evaporation of the solvent. A solid was obtained and analyzed by XRPD (X-Ray Powder Diffraction) and NMR. However, this analysis confirmed that the solid obtained was the diazoxide starting material and not a salt.

Salt preparation was carried out in non-aqueous media by dissolving diazoxide in anhydrous methanol in the presence of either sodium methoxide or potassium methoxide and stirring the mixture at 60° C. for 15 minutes. The mixture was then cooled to room temperature while stirred. After approximately two hours, a solid was recovered, isolated by filtration, and dried in vacuo. Analysis by XRPD confirmed that the solid obtained was the diazoxide starting material and not a salt.

4.5.2. Preparation of Salts in Methanol or Ethanol According to the '573 patent

The preparation of diazoxide salts in methanol and ethanol was attempted using 22 different counter-ions according to the methods described by the '573 patent. For example, 20 mg of diazoxide was dissolved in 5 mL of ethanol and stirred and heated to ensure dissolution of the diazoxide. To the stirred solution was added approximately 1 molar equivalents of sodium methoxide. The solution was stirred for approximately 10-15 minutes at 60° C., and cooled to room temperature for approximately 2 hours. The resulting solid precipitate was concentrated under a nitrogen stream, and collected by filtration. The product was dried in vacuo and analyzed by XRPD. As shown in FIG. 15, XRPD of the solids collected from the sodium methoxide experiment, (as well as the solids collected from the potassium methoxide experiment, which was run under similar conditions), revealed that the solid was the diazoxide starting material and that no sodium or potassium salt was prepared. In FIG. 15, (d) is the XRPD pattern of free form diazoxide, (b) is the XRPD pattern of the product of potassium methoxide in methanol, and (c) is the XRPD pattern of the product of sodium methoxide in methanol.

Although not wishing to be bound by any theory, it is believed that a possible explanation for the failure to prepare diazoxide salts by the methods of the '573 patent (i.e., in the presence of alcohols, e.g., methanol or ethanol), is that the alcohol may have an effect on the stability of the alkali salts of diazoxide. This was supported by UV spectroscopy analysis of alkali salts of diazoxide (sodium and potassium). Both the sodium and potassium salts of diazoxide were synthesized by reacting diazoxide with NaOH or KOH in MEK. Elemental analysis, NMR and XRPD confirmed that the salts were made.

Upon dissolution of the sodium or the potassium salt in acetonitrile, the UV spectrum shows a shift into the red region of the spectrum for the λmax from approximately 268 nm for the free form of diazoxide to approximately 298 nm for both the potassium and sodium salt (see FIG. 1). Similarly, these salts can be stabilized in aqueous solutions by elevating the pH to above 9.0. A similar shift in the absorption maximum at pH 9.0 is measured using UV spectroscopy. Subsequent adjustment of the pH from greater than 9.0 to less than 6.2 results in the hydrolysis of the salt as measured by the recovery of the UV absorption pattern of the diazoxide free base (see FIG. 2). In contrast, when the sodium or the potassium salt is dissolved in methanol, the UV-Vis spectrum of the salt was identical to that of the diazoxide starting material, (see FIG. 3).

These results demonstrate that using methanol as the solvent is incompatible with the synthesis of alkali salts of diazoxide. In addition, the results show that isolation of an alkali metal salt of diazoxide (or any other salt) in the presence of an alcohol, such as methanol, may not be possible.

4.5.3. Failure to Obtain Salts from Acidic Counter Ions

Salt formation with diazoxide was also attempted using acidic counter-ions, such as, for example, hydrochloric acid, maleic acid, sulfuric acid, phosphoric acid, sulfamic acid, acetic acid, citric acid, lactic acid, tartaric acid, malonic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclohexylsulfamic acid, fumaric acid, benzoic acid, undecylenic acid, salicylic acid and quinic acid. However, no salt formation was observed for any acid in any solvent.

For example, 100 mg of diazoxide was dissolved in 50 mL of acetone and heated to 35° C. To a stirred solution was added approximately 4.65 molar equivalents of HCl. The reaction mixture was allowed to cool to room temperature for approximately 3 hours, with no precipitation observed. Solvent was removed in vacuo. The resulting solid was analyzed by XRPD, and the observed XRPD pattern was consistent with the free form diazoxide starting material. In all cases, the attempted synthesis of diazoxide salts from acids was unsuccessful in all solvents attempted.

4.6. Preparation of Salts of Compounds of Formulae V-VIII

The chloride salt of 3-amino-4-methyl-1,2,4-benzothiadiazine-1,1-dioxide is prepared dissolving approximately 300 mg (1.4 mmol) in 45 mL acetonitrile. The mixture is heated to approximately 75° C. and stirred for 30 min. To the stirred solution, approximately 1 molar equivalent of HCl is added dropwise, and stirred for approximately 30 min at 75° C. The mixture is cooled to room temperature and the solvent is removed under reduced pressure, affording the chloride salt as a solid.

The sodium salt of 3-amino-4-methyl-1,2,4-benzothiadiazine-1,1-dioxide is prepared dissolving approximately 300 mg (1.4 mmol) in 45 mL acetonitrile. The mixture is heated to approximately 75° C. and stirred for 30 min. To the stirred solution, approximately 1 molar equivalent of NaOH is added dropwise, and stirred for approximately 30 min at 75° C. The mixture is cooled to room temperature and the solvent is removed under reduced pressure, affording the sodium salt as a solid.

5. Characterization of Prepared Diazoxide Salts

Synthesis of the desired salts was confirmed by X-Ray Powder Diffraction (XRPD), UV-Vis spectroscopy, and NMR. All spectra were compared with the spectra of the free form diazoxide (i.e., not a salt). Differential scanning calorimetry (DSC), thermal gravimetric analysis (TGA), FTIR (Fourier transform infrared spectroscopy), NMR, UV-vis spectroscopy and moisture sorption analysis were also performed.

5.1. Experimental Procedures

DSC analysis were conducted with a Mettler 822 DSC, by measuring the amount of energy released by a sample, as the sample was heated from 30° C. to between 300-500° C. at a rate of 10° C./min. Typical applications of DSC analysis include determination of melting point temperature and the heat of melting; measurement of the glass transition temperature; curing and crystallization studies; and identification of phase transformations.

TGA measurements were conducted with a Mettler 851 SDTA/TGA, by measuring weight loss as a function of increasing temperature, as the samples were heated from 30° C. to 230° C. at a rate of 10° C./min. The TGA can be used to analyze desorption and decomposition behavior, characterize oxidation behavior, set burnout or conditioning parameters (temperature/ramp rate/time), and determine chemical composition.

XRPD samples were analyzed with a Shimadzu XRD-6000 system, using a Cu Kα, 40 kV, 40 mA X-ray tube. The divergence and scatter slits were 1.00 deg, and the receiving slit was 0.30 mm. Samples were continuously scanned at a range of 3.0-45.0 deg, with a step size of 0.04 deg., at a scan rate of 2 deg/min.

Fourier Transform Infrared Spectroscopy was measured with a Thermo-Nicolet Avatar 370 with a Smart Endurance Attenuated Total Reflection (ATR) attachment. Compressed samples were analyzed, with corrections for background noise being made. Using the IR spectrum, chemical bonds and the molecular structure of organic compounds can be identified. Attenuated total reflectance (ATR) allows for the analysis of thin films, organic and inorganic, in areas as small as 10-15 microns.

Nuclear Magnetic Resonance (NMR) was performed with a 400 MHz Bruker Avance with a 4 mm CP/MAS H-X probe. Acquisition of 1H NMR spectra were performed by taking between 5-10 mg of the sample, dissolved in approximately 0.78 mL of DMSO-d6. Spectra were acquired with either 16 or 32 scans, using a pulse delay of 1.0 sec, with a 10 μsec(30°) pulse width.

UV spectroscopy was performed with a Perkin-Elmer Lambda 25 spectrometer. Samples were dissolved in acetonitrile, water and a buffer system having a pH between 5.6 and 10. Spectra were acquired between 340 and 190 nm, using a 1 cm path length with background correction.

Moisture Sorption Analysis was performed with a Hiden IGAsorp Moisture Sorption Instrument. Samples were first dried at 0% relative humidity at 25° C. until an equilibrium weight was reached, or for a maximum of 4 hours. Samples were then subjected to an isothermal (25° C.) scan from 10-90% relative humidity in steps of 10%. The samples were allowed to equilibrate to an asymptotic weight at each point for a maximum of 4 hours. Following absorption, a desorption scan from 85% relative humidity (at 25° C.) was run in steps of −10%, again allowing a maximum of 4 hours for the samples to equilibrate. The resulting samples after desorption were dried at 80° C. for two hours and analyzed by XRPD.

5.2. Free Form Diazoxide Characterization

The free form of diazoxide was characterized by XRPD, differential scanning calorimetry (DSC), thermal gravimetric analysis (TGA), moisture sorption, 1H NMR, FTIR and UV-vis spectroscopy to provide a baseline for comparison with the salts. Free form diazoxide is highly crystalline, as shown by the XRPD pattern. (See FIG. 4( a)). The DSC shows a large endothermic event at 330° C., and TGA shows that the free form of diazoxide is anhydrous, where diazoxide shows no weight loss below 200° C., and a weight loss of only 0.2% below 230° C. Moisture absorption of the free form diazoxide shows the material to be non-hygroscopic. Absorption of water by diazoxide was tested at between 0-90% relative humidity (RH) at 25° C., showing absorption of approximately 0.04 wt % at 60% RH and 0.20 wt % at 90% RH. The molecule does not form a stable hydrate, as shown by the lack of hysteresis during desorption. Additionally, the XRPD pattern for the diazoxide before and after absorption of water indicate the same crystalline form.

UV-vis spectroscopy measurements taken of the free form diazoxide in neutral aqueous solution show a λmax at approximately 268 nm. In acetonitrile, the λmax was 264 nm, demonstrating a small solvatochromic shift. As shown in FIG. 2, as pH increased the λmax also increased, from approximately 265 nm to approximately 280 nm, due to a change in the electronics of the molecule.

Studies were also conducted to evaluate the likelihood for conversion and degradation under thermal stress. Samples were heated in a closed environment, protected from light, at 60° C. for approximately 14 days. The diazoxide showed no conversion or degradation at 7 days or 14 days. Diazoxide samples were found to be consistent with the starting material with respect to XRPD and DSC.

Slurry studies to determine propensity of inter-conversion of the solid form were conducted on the free form diazoxide at room temperature, in the absence of light, using water, isopropyl alcohol, dichloromethane and toluene. Approximately 20 mg of the free form diazoxide was stirred for 14 days. Analysis by XRPD, DSC and HPLC were consistent with the starting material, indicating that the free form of diazoxide did not convert to alternate crystal forms.

5.3. Characterization of Sodium Diazoxide Salt

The XRPD pattern of the sodium salt of diazoxide was analyzed, showing the material to be crystalline. (See FIG. 4( d)). The DSC analysis revealed a major exothermic event at 448° C. Small transitions below 400° C. are likely due to sample imperfections. TGA analysis showed weight loss of 0.2% and 0.03% below 120° C., which may be the result of bound solvent. Moisture absorption performed from 0-90% relative humidity at 25° C. showed the material to be hygroscopic as the sample deliquesced at 90% relative humidity. The sample absorbed 1.2 wt % of water at 60% RH, and 6.6 wt % water at 80% RH. Hysteresis was observed upon desorption at 65 and 55% RH, indicating possible hydrate formation. (Possible hydrate formation was noted, although the amount of water absorbed was less than 0.5 mole). 1H NMR showed a chemical shift in the aromatic and methyl resonances of the sodium salt, as expected due to changes to the aromatic system. See FIG. 5 showing NMR spectrum for the free form of diazoxide (a) and the sodium salt of diazoxide (c). FTIR showed expected changes for the sodium salt.

Elemental analysis of the salt indicated that the salt was formed in a ratio of approximately 1:1, with the percentage of sodium being slightly low (approximately 3.4%). This deficiency may be due to matrix effects, as NMR indicated the sample had a relatively high purity.

UV-vis measurements in neutral aqueous solution show a λmax of approximately 271 nm (See FIG. 1). This value is slightly higher than free form diazoxide (265 nm). In acetonitrile, the λmax of the sodium salt exhibits a solvatochromic shift to approximately 296 nm. (See FIG. 3). An increase in the pH of the solution is expected to produce a bathochromic shift from approximately 265 nm to approximately 280 nm.

Solubility measurements performed at pH 2, 7, and 12 in 10 mM phosphate buffer at room temperature showed solubility of the sodium salt of diazoxide to be 13.0 mg/mL, 18.1 mg/mL and 48.6 mg/mL, respectively.

Form conversion and degradation under thermal stress were conducted as described for the free form diazoxide salt and showed the salt had little propensity to change form or degrade over a period of 14 days. Similarly, slurry studies were conducted as described for the free form diazoxide in n-heptane, dichloromethane and toluene showed no propensity of inter-conversion. See FIG. 6, wherein (a) is the XRPD pattern for the sodium salt of diazoxide, (b) is the XRPD pattern for the sodium salt after the slurry study, and (c) the XRPD of the free form of diazoxide.

5.4. Characterization of Potassium Diazoxide Salt

The XRPD pattern for the potassium salt of diazoxide was analyzed, showing the material to be crystalline. (See FIG. 4( b)). The DSC analysis revealed two major exothermic events at 128 and 354° C. (See FIG. 7). Small endotherms are likely due to sample impurities or the presence of solvent. TGA analysis showed weight loss of 7.7% below 220° C., which may be the result of moisture sorption. (See FIG. 8). Theoretical weight loss for a monohydrate of the diazoxide salt is 6.6%. Moisture absorption performed from 0-90% relative humidity at 25° C. showed the material to be hygroscopic as the sample deliquesced at 90% relative humidity, showing 38.3 wt % water. The sample absorbed 5.4 wt % of water at 60% RH. Hysteresis was observed upon desorption, however the material was determined to be a hemihydrate from 0-30% RH and a monohydrate from 35-75% RH. XRPD following the desorption analysis indicated that the sample had changed to an alternate crystalline form. 1H NMR showed a chemical shift in the aromatic and methyl resonances of the sodium salt, as expected due to changes to the aromatic system. (See FIG. 5 showing spectrum of the free form diazoxide (a) and the Potassium salt of diazoxide (b)). FTIR showed expected changes for the potassium salt.

Elemental analysis of the potassium salt indicated that the salt was formed in a ratio of approximately 1:1, with the percentage of potassium being slightly low (approximately 1.6%). This deficiency may be due to matrix effects, as NMR indicated the sample had a relatively high purity.

UV-vis measurements of the potassium diazoxide salt in neutral aqueous solution show a λmax of approximately 265 nm, which is equivalent to the diazoxide free form λmax. (See FIG. 1. In acetonitrile, the λmax of the potassium salt exhibits a solvatochromic shift to approximately 296 nm. (See FIG. 3). The potassium salt was used in a pH dependency study and showed that increasing the pH of the solution resulted in a bathochromic shift of the λmax from approximately 265 nm to approximately 280 nm.

Solubility measurements performed at pH 2, 7, and 12 in 10 mM phosphate buffer at room temperature showed solubility of the sodium salt of diazoxide to be 9.9 mg/mL, 14.4 mg/mL and 43.0 mg/mL, respectively. The potassium salt displayed greater solubility than the free form diazoxide, and demonstrated similar solubility to the sodium diazoxide salt. The XRPD pattern of solids obtained after the solubility analysis indicated that the potassium salt had changed back to the free form diazoxide material.

Propensity for form conversion and degradation under thermal stress were conducted as described for the free form diazoxide salt. The XRPD pattern of the sample after 7 and 14 days showed unique peaks, as compared with the potassium salt starting material. Analysis by DSC after 14 days also showed unique peaks as well. Using a gradient area percent assay, HPLC did not show any significant degradation of the potassium salt.

Slurry studies were conducted as described for the free form diazoxide in n-heptane, dichloromethane and toluene showed no propensity of inter-conversion. XRPD analysis of samples after 7 and 14 days showed unique peaks similar to those observed with the thermal stress study. See FIG. 9, wherein (a) is the XRPD pattern of the potassium salt of diazoxide, (b) is the XRPD pattern of the potassium salt of diazoxide after the slurry study in toluene, and (c) the XRPD pattern of the potassium salt after 14 days of the slurry study in toluene. Analysis by DSC after 14 days also showed unique peaks as compared with the starting material. HPLC using a gradient area percent assay did not show any significant degradation after the study.

5.5. Characterization of Choline Diazoxide Salt

The XRPD pattern of the choline salt of diazoxide was analyzed, showing the material to be crystalline. (See FIG. 10 (b)). The DSC analysis revealed a major exothermic events at 167° C. (See FIG. 11). A smaller endothermic event was seen at 119° C. and is likely due to sample impurities or the presence of residual solvent. TGA analysis showed weight loss of 0.8% between 100 and 140° C., which may be the result of residual solvents. (See FIG. 12). Moisture absorption performed from 0-90% relative humidity at 25° C. showed the material to be hygroscopic as the sample absorbed over 28% at 80% relative humidity, and deliquesced at 90% RH. Hysteresis was not observed upon desorption, indicating the choline salt does not form a stable hydrate. The XRPD pattern following the desorption analysis indicated that the sample had changed to an alternate crystalline form during the hydration and subsequent desorption. (See FIG. 13 (c)) 1H NMR was consistent with a 1:1 molar ratio of diazoxide to counter-ion, and had the expected differences in the chemical shift of the aromatic and methyl resonances, as expected due to the change in the local environment of the aromatic system due to the presence of the choline counter-ion. (See FIG. 14 (b)). FTIR showed expected changes for the choline salt, similar to those seen with the sodium and potassium salts.

Elemental analysis of the choline salt indicated that the salt was formed in a ratio of approximately 1:1. This is consistent with the 1H NMR.

UV-vis measurements of the choline diazoxide salt in neutral aqueous solution show a λmax of approximately 268 nm, which is close to the λmax for the diazoxide free form of 265 nm. In acetonitrile, the λmax of the choline salt exhibits a solvatochromic shift to approximately 296 nm, which is consistent with the sodium and potassium diazoxide salts. The potassium salt was used in a pH dependency study and showed that increasing the pH of the solution resulted in a bathochromic shift of the λmax from approximately 265 nm to approximately 280 nm.

Solubility measurements performed at pH 2, 7, and 12 in 10 mM phosphate buffer at room temperature showed solubility of the sodium salt of diazoxide to be 28.2 mg/mL, 41.5 mg/mL and greater than 293 mg/mL, respectively. The choline salt displayed greater solubility than the free form diazoxide after being allowed to equilibrate for 12 hours. The XRPD pattern of the solids obtained after the solubility analysis (at pH 2 and 7 only) indicated that the choline salt had changed back to the free form diazoxide material.

Propensity of the choline salt for form conversion and degradation under thermal stress were conducted as described for the free form diazoxide salt. XRPD analysis of the sample after 7 and 14 days showed an XRPD pattern consistent with the starting material, as well as the presence of additional unique peaks. (More unique peaks were present after 14 days than after 7 days). Analysis by DSC after 14 days did not show any significant difference, with a small endotherm at 117° C. and a large endotherm at 168° C. (Initial DSC showed endotherms at 119° C. and a large endotherm at 167° C.). Using a gradient area percent assay, HPLC did not show any significant degradation of the choline salt.

Slurry studies were conducted as described for the free form diazoxide in n-heptane, dichloromethane and toluene showed no propensity of inter-conversion. XRPD analysis of samples after 7 and 14 days showed signal associated with the starting material, with additional unique peaks present. (See FIG. 13 (c)). The XRPD pattern of slurry study samples from n-heptane showed signals associated with the starting material, as well as other additional unique signals. The XRPD pattern of slurry samples from dichloromethane and toluene were consistent with spectra obtained after the thermal studies and the moisture sorption analysis. Analysis by DSC after 14 days revealed a small endotherm at 109° C., and a major endotherm at 167° C. HPLC using a gradient area percent assay did not show any significant degradation after the study.

5.6. Characterization of Diazoxide Salt of Hexamethyl Hexamethylene Diammonium Hydroxide (HHDADH)

The XRPD pattern of the HHDADH salt of diazoxide was analyzed, showing the material to be a crystalline solid. (See FIG. 10( c)). Integration of the 1H NMR spectra was consistent with a 2:1 molar ratio of diazoxide to counter-ion (wherein the HHDADH counter-ion is divalent), and had the expected differences in the chemical shift of the aromatic and methyl resonances due to the change in the local environment of the aromatic system due to the presence of the choline counter-ion. (See FIG. 14( c)). The λmax of the HHDADH diazoxide salt in acetonitrile measured by UV-vis is 296 nm, which is consistent with the sodium, potassium and choline diazoxide salts.

A summary of the characterization of the free form diazoxide and the potassium, sodium, choline and hexamethyl hexamethylene diammonium hydroxide salts of diazoxide are presented in Table 14.

TABLE 14 Summary of Characterization and Solubility for Diazoxide and Salts Potassium Sodium Choline Test Free form diazoxide salt diazoxide salt diazoxide salt HHDADH XRPD crystalline crystalline crystalline crystalline crystalline FTIR consistent consistent consistent consistent N/A DSC single multiple Single two N/A endotherm at endotherms below exotherm endotherms 330° C. 300° C. above 400° C. below 200° C. UV in 268 nm 265 nm 271 nm 268 nm N/A aqueous 264 nm 296 nm 296 nm 296 nm 296 nm acetonitrile TGA <1% 7.7 (monohydrate <1% <1% N/A 6.7%) Moisture non- deliquescent - deliquescent - deliquescent - N/A Sorption hygroscopic - hemi/monohydrate potential no hysteresis no hysteresis hemihydrate Solubility pH 2.0 0.4 mg/mL 9.9 mg/mL 13.0 mg/mL 28.2 mg/mL N/A pH 7.0 0.04 mg/mL 14.4 mg/mL 18.1 mg/mL 41.5 mg/mL pH 12.0 4.8 mg/mL 43.0 mg/mL 48.6 mg/mL >293 mg/mL

Additional solubility studies were conducted for diazoxide free base form and diazoxide choline salt, as reported in Table 15. Each determination was carried out in duplicate, slurrying each sample in a 100 mM phosphate buffer solution, pH 7.00. Duplicate samples were then titrated to pH 7.0 and pH 8.8 using a 0.1 N phosphoric acid solution, followed by stirring for 18 h at ambient temperature. After this time all samples were centrifuged, and the supernatant was diluted with mobile phase (MeCN/Water.). Solubility was then obtained using a calibration curve for diazoxide by HPLC analysis. In Table 15, the term “Diazoxide, free from” refers to the free base of diazoxide; the term “Diazoxide, choline salt” is the choline salt of diazoxide as described herein; the term “Diazoxide, choline salt, milled” refers to the choline salt of diazoxide which has been milled by methods described herein. Table 15 shows that in 100 mM phosphate buffer, pH 7, and with titration with 0.1N phosphoric acid to a pH of about 6.8 to 8.8, solubility is notably suppressed compared to solubility at pH 10-11. Furthermore, the diazoxide choline salts were found to have increased solubility when compared to the parent free base.

TABLE 15 Summary of Solubility for Diazoxide and Diazoxide Choline Salt. pH after Phosphate titration pH Sample buffer 1N with after amount amount H₃PO₄ 0.1N 18 h Solubility Sample (mg) (mL) (mL) H₃PO₄ slurry [mg/mL] Diazoxide, 49.23 1 0.01 6.87 6.90 0.07 free form Diazoxide 47.64 1 — 7.30 7.31 0.07 free form Diazoxide, 44.34 1 0.15 7.22 7.45 0.12 choline salt Diazoxide, 48.26 1 0.11 8.81 8.64 0.18 choline salt Diazoxide, 45.35 1 0.14 7.32 7.36 0.12 choline salt, milled Diazoxide, 53.15 1 0.16 8.50 8.52 0.20 choline salt, milled Diazoxide, 50.0 1 — — 10.57 42.11 choline salt Diazoxide, 50.0 3 — — 10.52 32.88 choline salt, milled “—” Indicates no titration was conducted.

6. Polymorphic Forms of Diazoxide Salts

Polymorphic forms of the salts of diazoxide, characterization, and preparation thereof are described.

6.1. Polymorphic Forms of the Choline Salt of Diazoxide

6.1.1. Demonstration of Preparation of Polymorphic Form B of the Choline Salt of Diazoxide

A 1-L round bottom flask was charged with diazoxide (5 g), MEK (750 mL) and choline hydroxide (5.25 g of 50 wt % solution in water), and heated to 77° C. The mixture was allowed to cool to approximately ˜30° C., and filtered to remove insoluble brown residues. The filtrate was then concentrated under reduced pressure to afford yellow oil, which was dried in vacuo at 55° C. and 30 in·Hg to afford approximately 7.8 g as a waxy solid. The solids were dried in vacuo at 55° C. and 30 in·Hg to afford 7.13 g of the choline salt of diazoxide as a crystalline solid. Elemental analysis. Theoretical: C 46.77%, H 6.04% and N 12.59%. Measured: C 45.44%, H 5.98% and N 11.46%.

6.1.2. Characterization of the Polymorphic Form B of the Choline Salt of Diazoxide

The polymorphic Form B of the choline salt of diazoxide was analyzed by XRPD, DSC, and 1H NMR. As shown in FIG. 16, the two identified polymorphic forms of the choline salt of diazoxide show different XRPD patterns. See FIG. 16, wherein (a) shows the polymorphic Form A of the choline salt of diazoxide.

As shown in FIG. 17, 1H NMR of the polymorphic Form B of the choline salt of diazoxide shows no change from the polymorphic Form A.

Moisture absorption of the polymorphic Form B crystal structure of the choline salt of diazoxide performed from 0-80% relative humidity (RH) at 25° C. showed the material to be hygroscopic as the sample absorbed over 14.5% at 70% relative humidity, and deliquesced at 80% RH. The XRPD pattern following the desorption analysis indicated that the sample remained in the Form B crystal structure during the hydration and subsequent desorption.

Solubility measurements performed at pH 2, 7, and 12 in 10 mM phosphate buffer at room temperature showed solubility of the Form B crystal structure of the choline salt of diazoxide to be 32.8 mg/mL, 80.1 mg/mL and 216 mg/mL, respectively. The XRPD pattern of the solids obtained after the solubility analysis (at pH 2 and 7) indicated that Form B of the choline salt of diazoxide was still present.

Slurry experiments were performed on each form to determine the propensity for conversion and to search for possible new and/or unique forms. Upon slurrying Form B in CH₂Cl₂, n-heptane, and toluene, form conversion was not observed.

6.1.3. Demonstration of Preparation of the Polymorphic Form A of the Choline Salt of Diazoxide from the Polymorphic Form B of the Choline Salt of Diazoxide

Approximately 20 mg of the Form B polymorph of the choline salt of diazoxide was added to approximately 1 mL of acetone and heated to approximately 55° C. The mixture was filtered while hot and placed in a refrigerator (4° C.) for 16 hours. No precipitate was observed. The solvent was evaporated down to dryness using a gentle stream of nitrogen. The resultant solids were also dried in vacuo at room temperature and 30 in·Hg. XRPD analysis showed the salt had converted from the Form B polymorph to the Form A polymorph.

6.1.4. Characterization of the Polymorphic Form A of the Choline Salt of Diazoxide

Form A is an anhydrous crystalline form of diazoxide choline, with an endothermic event at approximately 165° C. in the DSC (see FIG. 20). The XRPD pattern for Form A is unique compared to that of Form B as shown in FIG. 21. FTIR (ATR) spectroscopy additionally indicates differences between the two forms. 1H NMR analysis affords a spectrum consistent with diazoxide and a 1:1 ratio of compound/counterion. NMR data also indicate that the magnetic environment of the diazoxide structure changes between free and polymorph forms, as evidenced by a movement in chemical shift of the aromatic and methyl proton resonances. In addition, the resonance due to the amine proton is not observed which suggests deprotonation in solution. Weight loss by TGA is less than 1% and may be due to residual solvent. The temperature of weight loss is above 100° C. which suggests that solvent may have been bound (i.e., solvate material). Moisture-sorption analysis conducted at 25° C. from 0 to 80% RH (adsorption) and 75 to 0% RH (desorption) shows Form A to be a hygroscopic solid, showing 2.4 wt % water at 60% RH. The sample was found to have deliquesced above 75% RH. In comparison, Form B is also hygroscopic and showed 7.4 wt % water at 60% RH and deliquesced at 80% RH. XRPD analysis following the moisture-sorption experiment affords a pattern consistent with Form A. Solubility studies conducted on both forms at pH 2, 7, and 12 in phosphate buffer showed differences, with Form A showing 28, 41, and >293 mg/mL respectively. Solubility concentrations were determined using area-percent calculations with HPLC calibration curves. Slurry experiments were performed on each form to determine propensity for conversion and to see if a unique form could be generated. Upon slurrying Form A in CH₂Cl₂, n-heptane, and toluene, conversion to Form B was observed after seven days. These results suggest that Form A is less thermodynamically stable under these conditions than Form B according to Ostwald's Rule of Stages.

6.1.5. Screening for Polymorphic Forms of Diazoxide Choline Salt.

A polymorph screening study of diazoxide choline salt was conducted with a series of crystallization and slurry conditions. As described herein, interconversion of diazoxide choline salt forms A and B was observed during this investigation. Each polymorphic form of diazoxide choline salt resulting from this study was characterized using the techniques and procedures described herein. A summary of characterization tests is listed in Table 16.

TABLE 16 Characterization of Forms A and B of Diazoxide Choline Salt In Screening Study Experiment Form A Form B XRPD* Crystalline Crystalline DSC Endotherm ≈165° C. Endotherm ≈160° C. TGA <1% ≈1% FTIR (ATR)** Consistent w/structure Consistent w/structure ¹H NMR Consistent w/1:1 ratio Consistent w/1:1 ratio Moisture Hygroscopic - deliquesced Hygroscopic - deliquesced Sorption at 90% RH at 80% RH Solubility pH 2: 28 mg/mL pH 2: 33 mg/mL pH 7: 41 mg/mL pH 7: 80 mg/mL pH 12: >293 mg/mL pH 12: 216 mg/mL Thermal Started to convert to Form Stable after 14 days at B after 14 days at 60° C. 60° C. Slurries Converted to Form B after Stable after 14 days in 7 days in n-heptane, CH2Cl2, THF, and toluene, and CH2Cl2 t-AmOH *Major peaks (2-): Form A (9.8, 10.5, 14.9, 17.8, 17.9, 18.5, 19.5, 22.1, 22.6, 26.2, 29.6, 31.2); Form B (8.9, 10.3, 12.0, 18.3, 20.6, 24.1, 24.5, 26.3, 27.1, 28.9). **Unique FTIR (ATR) absorbances (cm⁻¹): Form A (2926, 2654, 1592, 1449, 1248); Form B (3256, 2174, 2890, 1605, 1463, 1235).

6.1.5.1. Solubility Screen in Organic Solvents.

Diazoxide choline, prepared in MEK using choline hydroxide as 50 wt % solution in water (see above) displayed some solubility in the following solvents: acetonitrile, acetone, ethanol, IPA, MEK, DMF, and methanol. These solvents were chosen due to differences in functionality, polarity, and boiling points and their ability to dissolve diazoxide. Other solvents which showed poor ability to dissolve salts were used as antisolvents and in slurry experiments where some solubility was observed: dioxane, MTBE, EtOAc, IPAc, THF, water, cyclohexane, heptane, CH₂Cl₂, and toluene.

Solvents for crystallizations during screening were chosen based on the solubility screen summarized in Table 17. Crystallizations of diazoxide choline from all conditions afforded a total of two forms, A and B. Forms A and B were found to be anhydrous polymorphs of diazoxide choline. Form B was observed to be generated from most solvents used. It was difficult to isolate pure Form A on large scales (>50 mg) as conditions observed to produce Form A on a smaller scale (approximately 50 mg or less) were found to result in Form B or mixtures of both forms on larger scales. Based on room-temperature slurry experiments, anhydrous Form B was found to be the most thermodynamically stable form in this study. Form A readily converted to Form B in all slurry solvents utilized.

TABLE 17 Solubility Screen for Diazoxide Choline Salt Cmpd Solvent Conc. Temp. Solvent (mg) (mL) (mg/mL) (° C.) Soluble MeCN 1.7 0.25 >6.80 rt Yes Dioxane 1.4 5.00 0.28 55 No Acetone 1.9 0.25 7.60 55 Yes MTBE 2.4 5.00 0.48 55 No EtOH 1.5 0.25 >6.00 rt Yes EtOAc 1.2 5.00 0.24 55 No IPAc 1.4 5.00 0.28 55 No IPA 1.8 0.25 7.20 55 Yes THF 1.1 5.00 0.22 55 No MEK 1.8 1.00 1.80 55 Yes DMF 1.2 0.25 >4.80 rt Yes Water 2.0 5.00 0.40 55 No MeOH 1.9 0.25 7.60 55 Yes c-Hexane 2.0 5.00 0.40 55 No Heptane 1.9 5.00 0.38 55 No CH₂Cl₂ 1.3 5.00 0.26 55 Partially Toluene 1.4 5.00 0.28 55 No

6.1.5.2. Single-Solvent Crystallizations

Fast cooling procedure: Diazoxide (approximately 20 mg) was weighed out into vials and enough solvent (starting with 0.25 mL) was added until the material completely dissolved at elevated temperature. After hot filtration the vials were placed in a refrigerator (4° C.) for 16 hours. After the cooling-process the samples were observed for precipitates which were isolated by filtration. Vials not demonstrating precipitates were evaporated down to dryness using a gentle stream of nitrogen. All solids were dried in vacuo at ambient temperature and 30 in·Hg.

Slow cooling procedure: Diazoxide (approximately 30 mg of choline salt) was weighed out into vials and enough solvent was added until the material went into solution at elevated temperature. After hot filtration the vials were then slowly cooled to room temperature at the rate of 20° C./h and stirred at room temperature for 1-2 hours. All solids were dried in vacuo at ambient temperature and 30 in·Hg.

Based on the initial solubility study, seven solvents were selected for the fast-cooling crystallization: acetonitrile, acetone, ethanol, IPA, MEK, DMF, and methanol. Table 18 shows a list of the solvents that were used and the amount of solvent needed to dissolve the material. After the cooling-process precipitates were noticed in samples #2, 3, 5, and 6, the solids were isolated by filtration. The other samples (#1, 4, and 7) were evaporated down to dryness using a gentle stream of nitrogen. The diazoxide choline salts were found to be consistent with Form A by XRPD analysis for all solids with the exception of sample #2 (consistent with the freeform) and sample #5 (consistent with Form B with preferred orientation observed).

TABLE 18 Single-Solvent Crystallization of Diazoxide Choline Salt Using Fast- Cooling Procedure Solvent BP Cmpd Amt Conc Temp. Entry Solvent (° C.) (mg) (mL) (mg/mL) (° C.) Precipitate Form 1 Acetone 56 21.0 1.00 21.00 55 No/Evap A 2 MeOH 64 20.3 0.25 81.20 55 Yes FF* 3 EtOH 78 21.3 0.25 85.20 62 Yes A 4 MEK 80 19.6 1.25 15.68 75 No/Evap A 5 MeCN 81 20.6 0.25 82.40 55 Yes Unique 6 IPA 82 22.8 0.25 91.20 62 Yes A 7 DMF 153 26.0 0.25 104.00 55 No/Evap A

In accordance with the data obtained from fast-cooling experiments, four solvents which showed precipitation of solids were chosen for the slow-cooling experiments: MeOH, EtOH, MeCN, and IPA (Table 19). All obtained analyzable solids of the choline salt were found to be consistent with Form B by XRPD with the exception of Entry #1 which was consistent with diazoxide freeform and Entry #2 which was not analyzable. Mother liquor of Entry #2 was concentrated to dryness and the residual solids were analyzed by XRPD and found to be Form B material. As a result of obtaining freeform material from the single-solvent crystallizations in methanol, three more alcohols were tested for the single-solvent crystallizations using fast- and slow-cooling procedures. Tables 20 and 21 provide a list of the solvents that were used and the amount of solvent needed to dissolve the material. XRPD patterns of the fast-cooling procedure showed freeform of diazoxide from isobutanol, Form B from isoamyl alcohol, and Form A from tert-amyl alcohol compared to the slow-cooling procedure, which afforded Form B material from all three solvents.

TABLE 19 Single-Solvent Crystallization of Diazoxide Choline Salt Using Slow- Cooling Procedure Boiling Material Solvent Point Amount Amount Conc. Temp. Solvent (° C.) (mg) (mL) (mg/mL) (° C.) Precipitate Form MeOH 64 32.1 0.3 107.00 62 Yes FF* EtOH 78 33.3 0.3 111.00 75 Yes NA** MeCN 81 30.9 0.3 103.00 62 Yes B IPA 82 33.7 0.3 112.33 80 Yes B

TABLE 20 Single-Solvent Crystallization of Diazoxide Choline Salt Using Fast- Cooling Procedure Boiling Material Solvent Point Amount Amount Conc. Temp. Solvent (° C.) (mg) (mL) (mg/mL) (° C.) Precipitate Form i-BuOH 108 29.7 0.3 99.00 78 Yes (sm)* i-AmOH 130 29.6 0.3 98.67 82 Yes B t-AmOH 102 29.5 0.3 98.33 95 No/Evap A

TABLE 21 Single-Solvent Crystallization of Diazoxide Choline Salt Using Slow- Cooling Procedure Boiling Material Solvent Point Amount Amount Conc. Temp. Solvent (° C.) (mg) (mL) (mg/mL) (° C.) Precipitate Form i-BuOH 108 33.0 0.3 110.00 92 Yes B i-AmOH 130 28.2 0.3 94.00 92 Yes B t-AmOH 102 29.0 0.4 72.50 92 Yes B

The results of the choline salt single-solvent fast- and slow-cooling crystallizations (see Tables 19 to 21) indicated that Form A was more likely to be isolated with fast-cooling profiles and Form B with slow-cooling profiles.

6.1.5.3. Binary Solvent Crystallizations

Binary-solvent crystallizations of the choline salt were performed using four primary solvents (MeOH, EtOH, IPA, and MeCN) and nine cosolvents (MTBE, EtOAc, IPAc, THF, c-hexane, heptane, toluene, CH2Cl2, and dioxane) with a fast-cooling profile (supra). XRPD patterns showed that Form B was obtained from mixtures of MeOH with MTBE, EtOAc, IPAc, toluene, and dioxane. As shown in Table 22, Form A was obtained from mixtures of MeOH with THF and with CH2Cl2 after evaporating the solvent to dryness. The mixtures of MeOH with cyclohexane and heptane provided the freeform of diazoxide. All solids obtained from fast-cooling procedures with EtOH, IPA, and MeCN as primary solvents provided Form B material.

TABLE 22 Binary-Solvent Crystallizations of Choline Salt of Diazoxide Using Fast-Cooling Procedure and MeOH as a Primary Solvent Diazoxide MeOH* Amount Amt (mg) (mL) Antisolvent (mL) Precipitate Form 27.8 0.3 MTBE 1.4 Yes B 30.7 0.3 EtOAc 6.0 Yes B 32.0 0.3 IPAc 6.0 Yes B 31.9 0.3 THF 6.0 No/Evap to Dry A 29.5 0.3 c-Hexane 2.0 Yes (small) FF** 30.2 0.3 Heptane 2.0 Yes FF** 29.3 0.3 Toluene 6.0 Yes B 32.0 0.3 CH2Cl2 6.0 No/Evap to Dry A 28.8 0.3 Dioxane 6.0 Yes B *Solids were dissolved at 62° C. **Freeform of diazoxide.

Binary-solvent recrystallizations of the choline salt with the slow-cooling procedure were performed using two primary solvents (IPA and MeCN) and nine cosolvents (MTBE, EtOAc, IPAc, THF, c-hexane, heptane, toluene, CH₂Cl₂, and dioxane). All solids obtained from a slow-cooling procedure with IPA and MeCN as primary solvents provided Form B material based on XRPD analysis. The results of binary-solvent crystallizations indicated that Form B was the most thermodynamically stable form of diazoxide choline.

6.1.5.4. Binary Solvent Crystallizations Using Water as a Cosolvent

In an attempt to investigate the formation of hydrates of the choline salt, experiments was performed using fast- and slow-cooling procedures and water as a cosolvent.

The fast cooling procedure (supra) was used with the exception of using different primary solvents which were miscible with water: acetone, acetonitrile, DMF, IPA, i-BuOH, i-AmOH, and t-AmOH. Water was utilized in these crystallizations as a cosolvent. All solids obtained from the fast-cooling procedure with water as the cosolvent provided diazoxide freeform material by XRPD analysis.

To compare the results obtained from the fast-cooling procedure a set of experiments was performed using a slow-cooling procedure and water as a cosolvent. All obtained solids were analyzed by XRPD and afforded patterns consistent with diazoxide freeform. Without wishing to be bound by theory, these results suggest that the conditions used for crystallization caused dissociation of the choline salt. A small amount of a second crop was obtained in each sample, but only two samples were analyzable by XRPD and indicated that the samples were freeform material. All mother liquors were evaporated to dryness and the residual solids were also analyzed by XRPD to afford patterns consistent with Form B of the choline salt.

6.1.5.5. Metastable Zone Width Estimation

Form B: To produce a robust process, an understanding of the solubility profiles of the various solid forms under consideration is required. From a practical standpoint, this involves the measurement of the metastable zone width (MSZW) of pure forms, whereby the saturation and supersaturation curves of the different forms are generated over a well defined concentration and temperature range. This knowledge can then be used to design a crystallization protocol that should ideally favor a selective crystal growth of the desired form.

Form B of diazoxide choline salt showed moderate solubility in a solvent mixture made of MeCN/MeOH/MtBE (10:1:12, volume ratios). The wide width of the metastable zone as shown in Table 23 gives many seeding options. During the MSZW measurement, aliquots from the crystallizing material were withdrawn and analyzed by XRPD to ensure that no form conversion occurred during the experiment. Indeed, the material remained unchanged during the test.

TABLE 23 Meta-Stable Zone Width For Form B Diazoxide Choline Salt in MeCN/MeOH/MtBE (10:1:12) (v/v). Conc.| Temp. In Temp. Out Temp. Range (mg/mL) (° C.) (° C.) (° C.) 30.8 53.2 35.0 18.2 28.5 49.0 33.6 15.4 26.5 47.0 32.0 15.0 24.7 43.8 29.1 14.7 23.2 40.5 28.5 12.0 21.9 38.0 26.0 12.0

Form A: The metastable zone width for Form could not be estimated because this polymorphic form converted during the experiment to Form B.

6.1.5.6. Crystallization of Form A of Diazoxide Choline Salt

The choline salt of diazoxide (160.3 mg) was dissolved in 1 mL of IPA at 55° C. which was then passed through a Millipore 0.45 μM filter into a clean vial. This vial was placed in freezer a −20° C. overnight. Solids were not noticed and the flask was scratched with a micro-spatula. The vial was placed back in the freezer and nucleation was noticed after ten minutes. The solids were collected by vacuum filtration and washed with 1 mL of MtBE. The solids were dried in vacuo at 40° C. and 30 in·Hg to afford 70 mg (43.6% recovery) of Form A as determined by XRPD.

6.1.5.7. 500-mg Scale Crystallization of Form B of Diazoxide Choline Salt

The choline salt of diazoxide (524.3 mg) was dissolved in 3 mL of IPA at 78° C. and this solution was then cooled to 55° C. for the addition of MtBE. The MtBE (4 mL) was added until nucleation was observed. After nucleation the batch was allowed to cool to room temperature at a rate of 20° C./h. The solids were collected by vacuum filtration and washed with 1 mL of MtBE. The solids were dried in vacuo at 40° C. and 30 in. of Hg to afford 426.7 mg (81.3% recovery) of Form B as determined by XRPD.

6.1.5.8. 2-g Scale Crystallization of Form B of Diazoxide Choline Salt

The choline salt of diazoxide (2.0015 g) was dissolved in 5.5 mL of IPA at 78° C. to afford a clear solution. This solution was passed through a Millipore Millex FH 0.45 μM filter. This solution was then cooled to 55° C. MtBE was added in 1 mL portions, with a two minute interval between portions. Nucleation was noted after the second addition of MtBE. This suspension was allowed to cool to room temperature at a rate of 20° C./h and stirred at this temperature for 16 hours. The solids were collected by vacuum filtration and washed with 1 mL of MtBE. The solids were dried in vacuo at 40° C. and 30 in. of Hg to afford 1.6091 g (80.4% recovery) of Form B as determined by XRPD.

6.1.5.9. Detection of Form Impurities

Mixtures of diazoxide choline Forms A and B were prepared by adding a minor amount of Form A to Form B. Samples were lightly ground by hands with a mortar and pestle for approximately one minute. Samples were then analyzed by XRPD analysis. XRPD analysis was found to be suitable for detecting 5% of Form A in Form B.

6.2. Polymorphic Forms of the Potassium Salt of Diazoxide

A summary of characterization tests for three common crystalline forms of diazoxide potassium salt are listed in Table 24. Solvents for crystallizations were chosen based on the solubility screen summarized below. Crystallizations of diazoxide potassium salt from all conditions provided herein afforded a total of seven unique crystalline forms, A through G. Forms C, D, and F were found to be the most common during the crystallization screen, and were therefore scaled up for further characterization.

TABLE 24 Results Summary of Characterization Tests for Salt of Diazoxide Potassium Experiment Form C Form D Form F XRPD* Crystalline Crystalline Crystalline DSC 187, 360° C. 130, 191, 352° C. 191, 363° C. TGA 8.4%** 4.5%** 13.1%** FTIR (ATR)* Consistent w/ Consistent w/ Consistent w/ structure structure structure ¹H NMR Consistent w/ Consistent w/ Consistent w/ structure** structure structure** Moisture Hygroscopic - Hygroscopic - Sorption deliquesced at 90% RH deliquesced at 90% RH Hygroscopic - deliquesced at 90% RH Solubility N/A pH 2: 29 mg/mL N/A pH 7: 33 mg/mL pH 12: 59 mg/mL Thermal Stable (7 days) Stable (7 days) Stable (7 days) Slurries Converted to Form D Converted to Form D Converted to Form D *XRPD Major peaks (2-): Form A (6.0, 8.1, 16.3, 17.7, 18.6, 19.1, 22.9, 23.3, 23.7, 24.7, 25.4, 26.1, 28.2, 29.6, 30.2); Form B (8.5, 10.8, 16.9, 18.2, 21.6, 25.5, 26.1, 28.9); Form C (5.7, 6.1, 17.9, 23.9, 25.1, 37.3); Form D (5.7, 6.2, 8.1, 8.5, 8.8, 16.9, 18.6, 23.2, 24.5, 25.8, 26.1); Form E (6.7, 7.1, 14.1, 21.2); Form F (8.5, 9.0, 18.7, 20.6, 23.5, 27.5, 36.3); Form G (5.2, 5.5, 13.1, 16.5, 19.3, 22.8, 24.8, 26.4, 28.7, 3 4.1); *Unique FTIR (ATR) absorbances (cm⁻¹): Form A (1503, 1374, 1339, 1207, 1131, 1056, 771); Form B (1509, 1464, 1378, 1347); Form C (1706, 1208, 1146, 746); Form D (1595, 1258, 1219, 890); Form E (1550, 1508, 1268, 1101, 1006). Form F (1643, 1595, 1234, 1145, 810). Form G (1675, 1591, 1504, 1458, 1432, 1266, 999, 958, 905, 872). **Data indicates a half-molar equivalent of acetone, water, or dioxane for Forms C, D, and F respectively.

Diazoxide potassium Forms C, D, and F were observed to be an acetone solvate, a hemihydrate, and a dioxane solvate of diazoxide potassium, respectively. Form C is an acetone solvate that was generated predominantly when acetone was used in the crystallization. Form D, a hemihydrate, was observed to be generated from most solvents used. Form F is a dioxane solvate generated when dioxane was used as an antisolvent. Forms A, B, E, and G were not commonly observed during the crystallization. Elemental analysis data indicated that the unique forms observed may be mixtures and/or have residual solvent(s) present.

Based on room-temperature slurry experiments presented, Form D was found to be the most thermodynamically stable form of those discovered in this study. Forms C and F readily converted to Form D in all slurry solvents utilized. Without wishing to be bound by theory, since nonaqueous solvents were used, the material may have converted to the hemihydrate, Form D, upon removal from the solvent.

6.2.1. Demonstration of Preparation of Polymorphic Form A of the Potassium Salt of Diazoxide

The polymorphic Form A of the potassium salt of diazoxide was prepared as described above.

6.2.2. Demonstration of Preparation of the Polymorphic Form B of the Potassium Salt of Diazoxide

Diazoxide (2.95 g) was combined with 450 mL of methyl ethyl ketone and heated to approximately 77° C. to dissolve the diazoxide. To the solution was added approximately 13 mL of 1M potassium hydroxide at a rate of approximately 20 mL/min, stirred and allowed to cool to room temperature. The solution was stirred at room temperature for ˜16 h. The solvent was removed under reduced pressure, and the residual solids were dried in vacuo at 57° C. and 30 in·Hg to afford 3.7 g of the potassium salt.

6.2.3. Characterization of the Polymorphic Form B of the Potassium Salt of Diazoxide

The Form B polymorph of the potassium salt of diazoxide was analyzed by XRPD, and 1H NMR. FIG. 18 shows the XRPD pattern of (a) the Form A polymorph of the potassium salt of diazoxide and (b) the Form B polymorph of the potassium salt of diazoxide.

The 1H NMR of the Form B polymorph of the potassium salt of diazoxide shows no change from the Form A polymorph.

6.2.4. Demonstration of Preparation of the Polymorphic Form A of the Potassium Salt of Diazoxide from Form B

Approximately 20 mg of the Form B polymorph of the diazoxide potassium salt was added to 2 mL of acetone and heated until the material completely dissolved at 55° C. The solution was hot filtered and placed in a refrigerator (4° C.) for 16 hours. No precipitate was formed. The solvent was evaporated down to dryness using a gentle stream of nitrogen and the resultant solids were dried in vacuo at room temperature and 30 in·Hg. The solid was analyzed by XRPD to determine the physical form. See FIG. 18( a).

6.2.5. Preparation of the Polymorphic Form C of the Potassium Salt of Diazoxide from Form B

Approximately 20 mg of the Form B polymorph of the diazoxide potassium salt was added to 6 mL of ethyl acetate and heated until the material completely dissolved at 75° C. The solution was hot filtered and placed in a refrigerator (4° C.) for 16 hours. No precipitate was formed. The solvent was evaporated down to dryness using a gentle stream of nitrogen and the resultant solids were dried in vacuo at room temperature and 30 in·Hg. The solid was analyzed by XRPD to determine the physical form. See FIG. 18( c).

6.2.6. Preparation of the Polymorphic Form D of the Potassium Salt of Diazoxide from Form B

Approximately 20 mg of the Form B polymorph of the diazoxide potassium salt was added to 0.3 mL of isopropyl alcohol and heated until the material completely dissolved at 62° C. The solution was hot filtered and placed in a refrigerator (4° C.) for 16 hours. No precipitate was formed. The solvent was evaporated down to dryness using a gentle stream of nitrogen and the resultant solids were dried in vacuo at room temperature and 30 in·Hg. The solid was analyzed by XRPD to determine the physical form. See FIG. 19( a).

6.2.7. Preparation of the Polymorphic Form E of the Potassium Salt of Diazoxide from Form B

Approximately 20 mg of the Form B polymorph of the diazoxide potassium salt was added to 2.5 mL of tert-amyl alcohol and heated until the material completely dissolved at 95° C. The solution was hot filtered and placed in a refrigerator (4° C.) for 16 hours. No precipitate was formed. The solvent was evaporated down to dryness using a gentle stream of nitrogen and the resultant solids were dried in vacuo at room temperature and 30 in·Hg. The solid was analyzed by XRPD to determine the physical form. See FIG. 19( b).

6.2.8. Preparation of the Polymorphic Form F of the Potassium Salt of Diazoxide from Form B

Approximately 20 mg of the Form B polymorph of the diazoxide potassium salt was added to 0.6 mL of acetonitrile and heated until the material completely dissolved at 80° C. The solution was hot filtered, 6 mL of dioxane was added, and the solution was placed in a refrigerator (4° C.) for 16 hours. No precipitate was formed. The solvent was evaporated down to dryness using a gentle stream of nitrogen and the resultant solids were dried in vacuo at room temperature and 30 in·Hg. The solid was analyzed by XRPD to determine the physical form. See FIG. 19( c).

6.2.9. Preparation of the Polymorphic Form G of the Potassium Salt of Diazoxide from Form B

Approximately 22.3 mg of the Form B polymorph of the diazoxide potassium salt was added to 0.5 mL of isoamyl alcohol and heated until the material completely dissolved at 73° C. The solution was hot filtered, 6 mL of isopropyl acetate was added, and the solution was placed in a refrigerator (4° C.) for 16 hours. No precipitate was formed. The solvent was evaporated down to dryness using a gentle stream of nitrogen and the resultant solids were dried in vacuo at room temperature and 30 in·Hg. The solid was analyzed by XRPD to determine the physical form. See FIG. 19( d).

As shown in Tables 25 and 26, both the solvent used for recrystallization and the rate of cooling (i.e., fast cooling vs. slow cooling during the recrystallization) effect the crystal structure obtained once the product is isolated.

TABLE 25 Fast cooling of Potassium Salt of Diazoxide in Various Solvents Cmpd Solvent Recovery Solvent (mg) (mL) (mg) Form THF 20.8 6.5 14.2 B EtOAc 21.1 6.0 10.6 C MeCN 21.2 0.5 n/a C + D IPA 22.7 0.3 n/a D Water 20.6 1.5 n/a Free form diazoxide tAA 21.9 2.5 16.7 E IAA 19.7 0.3 n/a D DMF 21.6 0.3 15.9 G

TABLE 26 Slow cooling of Potassium Salt of Diazoxide in Various Solvents Cmpd Solvent Recovery Solvent (mg) (mL) (mg) Form EtOAc 20.4 6.0 7.1 D MeCN 22.8 0.5 13.8 C IPA 21.5 0.3 13.2 C Water 20.9 1.5 1.2 Free form diazoxide tAA 21.5 2.5 15.4 E IAA 20.6 0.3 6.6 D

6.2.10. Polymorphs Obtained by Recrystallization of the Potassium Salt of Diazoxide in Binary Solvents

As shown in Tables 27 and 28, recrystallization of the potassium salt of diazoxide from a variety of binary solvent systems also demonstrated conversion of the potassium salt to an alternate form. Use of acetonitrile as the primary solvent is shown in Table 27 and use of acetone as the primary solvent is shown in Table 28. As shown in Table 27, recrystallization of the Form B polymorph of the potassium salt of diazoxide from acetonitrile using methyl tert-butyl ether, ethyl acetate, isopropyl acetate, tetrahydrofuran, c-hexane, heptane, toluene and dichloromethane as the secondary solvent all yielded the D Form polymorph of the potassium salt. Recrystallization from acetonitrile using dioxane as the secondary solvent yielded the F Form polymorph of the diazoxide salt of potassium.

TABLE 27 Recrystallization in Acetonitrile Acetonitrile Secondary Cmpd Recovery (mL) solvent (mg) (mg) Form 0.6 MTBE 20.7 13.7 D 0.6 EtOAc 23.4 9.5 D 0.6 IPAc 20.0 13.3 D 0.6 THF 20.3 6.4 D 0.6 c-Hexane 20.4 9.6 D 0.6 Heptane 20.3 10.8 D 0.6 Toluene 23.6 16.1 D 0.6 Dichloromethane 21.3 12.7 D 0.6 Dioxane 20.7 12.6 F

As shown in Table 28, recrystallization of the Form B polymorph of the potassium salt of diazoxide from acetone using methyl tert-butyl ether, tetrahydrofuran, and c-hexane as the secondary solvent all yielded the Form A polymorph of the potassium salt of diazoxide. Recrystallization from acetone using ethyl acetate, heptane, toluene and dichloromethane as the secondary solvent all yielded the C Form polymorph of the potassium salt. Recrystallization from acetone using isopropyl acetate as the secondary solvent yielded the D Form polymorph of the diazoxide salt of potassium. Recrystallization from acetone using dioxane as the secondary solvent yielded the F Form polymorph of the diazoxide salt of potassium.

TABLE 28 Recrystallization in Acetone Acetone Amt Recovery (mL) Other Solvent (mg) (mg) Form 2.0 MTBE 20.2 13.9 A 2.0 EtOAc 21.6 4.8 C 2.0 IPAc 20.6 11.6 D 2.0 THF 20.9 12.0 A 2.0 c-Hexane 21.3 12.3 A 2.0 Heptane 20.6 12.7 C 2.0 Toluene 20.4 13.3 C 2.0 Dichloromethane 21.1 13.0 C 2.0 Dioxane 20.4 12.5 F

6.2.11. Screening for Polymorphic Forms of Diazoxide Potassium Salt.

A polymorphic screening study of diazoxide potassium salt was conducted with a series of crystallization conditions described below.

6.2.11.1. Solubility Screening for Polymorphic Forms of Diazoxide Potassium Salt.

Diazoxide potassium, prepared in MEK using 1 M potassium hydroxide solution in water, displayed some solubility in the following ten solvents: acetone, THF, EtOAc, MEK, MeCN, IPA, water, t-AmOH, i-AmOH, and DMF. These solvents were chosen due to differences in functionality, polarity, and boiling points and their ability to dissolve diazoxide. Solvents affording poor to fair solubility were used as antisolvents in binary/ternary crystallizations as well as slurry studies. Table 29 summarizes the results of the solubility screen.

TABLE 29 Solubility of Diazoxide Potassium Salt in Various Solvent Cmpd Amt Solvent Conc. Temp. Solvent (mg) (mL) (mg/mL) (° C.) Soluble MeCN 1.7 2.00 0.85 55 Yes Dioxane 1.4 5.00 0.28 55 No Acetone 1.6 4.00 0.40 55 Yes MTBE 1.8 5.00 0.36 55 No EtOH 2.2 0.75 2.93 55 Yes EtOAc 1.8 5.00 0.36 55 No IPAc 1.7 5.00 0.34 55 No IPA 2.1 1.00 2.10 55 Yes THF 1.8 5.00 0.36 55 Partially MEK 1.5 5.00 0.30 55 Partially DMF 1.6 0.25 >6.40 rt Yes Water 1.5 5.00 0.30 55 No MeOH 1.5 0.25 6.00 55 Yes c-Hexane 1.5 5.00 0.30 55 No Heptane 1.2 5.00 0.24 55 No CH₂Cl₂ 1.3 5.00 0.26 55 No Toluene 1.4 5.00 0.28 55 No

6.2.11.2. Single-Solvent Screening for Polymorphic Forms of Diazoxide Potassium Salt.

Single-solvent crystallizations of potassium salt were performed using ten solvents: acetone, THF, EtOAc, MEK, MeCN, IPA, water, t-AmOH, i-AmOH, and DMF for the fast-cooling procedure and six solvents (EtOAc, MeCN, IPA, water, t-AmOH, and i-AmOH) for the slow-cooling procedures. The “fast” and “slow” cooling procedures were as described above. Four of the solvents were excluded from the slow-cooling experiments because they did not provide solids during fast-cooling experiments and needed to be evaporated to dryness. Tables 30 and 31 provide a list of the solvents that were used and the amount of solvent needed to dissolve the material. All solids were analyzed by XRPD to determine the physical form and six unique patterns (Forms A-E, G) were observed.

TABLE 30 Single-Solvent Crystallizations of Potassium Salt of Diazoxide Using Fast- Cooling Cmpd Solvent Conc BP Amt Amt (mg/ Temp Solvent (° C.) (mg) (mL) mL) (° C.) Precipitate Form Acetone 56 20.7 2.0 10.35 55 No/Evap A THF* 65 20.8 6.5 3.20 63 No/Evap B EtOAc 76 21.1 6.0 3.52 75 Yes C MEK 80 20.2 4.0 5.05 75 No/Evap A MeCN 81 21.2 0.5 42.40 80 Yes C + D IPA 82 22.7 0.3 75.67 62 Yes D Water 100 20.6 1.5 13.73 95 Yes FF t-AmOH 103 21.9 2.5 8.76 95 Yes E i-AmOH 130 19.7 0.3 65.67 73 Yes D DMF 153 21.6 0.3 72.00 RT No/Evap G *Solids were not completely dissolved.

TABLE 31 Single-Solvent Crystallizations of Potassium Salt of Diazoxide Using Slow- Cmpd Solvent Conc BP Amt Amt (mg/ Temp Solvent (° C.) (mg) (mL) mL) (° C.) Precipitate Form EtOAc 76 20.4 6.0 3.40 75 Yes D MeCN 81 22.8 0.5 45.60 80 Yes C IPA 82 21.5 0.3 71.67 80 No/Evap C Water 100 20.9 1.5 13.93 95 Yes FF t-AmOH 103 21.5 2.5 8.60 95 Yes E i-AmOH 130 20.6 0.3 68.67 80 Yes D

6.2.11.3. Binary-Solvent Screening for Polymorphic Forms of Diazoxide Potassium Salt.

Binary-solvent crystallizations of the potassium salt utilizing fast-cooling procedure were performed using MeCN, acetone, and isoamyl alcohol as primary solvents and the following nine cosolvents: MTBE, EtOAc, IPAc, THF, c-hexane, heptane, toluene, CH₂Cl₂, and dioxane. Table 32 is representative, employing acetonitrile as primary solvent. XRPD patterns from crystallizations using acetonitrile as a primary solvent were consistent with Form D with only one exception being the solids obtained from the mixture of MeCN/dioxane afforded a unique pattern (Form F) material.

TABLE 32 Binary-Solvent Crystallizations of Potassium Salt of Diazoxide Using Fast-cooling Procedure and McCN as a Primary Solvent Cmpd Amt McCN* Anti- Amt (mg) (mL) solvent (mL) Precipitate Form 20.7 0.6 MTBE 1.0 Yes D 23.4 0.6 EtOAc 6.0 Yes D 20.0 0.6 IPAc 3.0 Yes D 20.3 0.6 THF 6.0 No/Evap. D To ppt 20.4 0.6 c-Hexane 2.0 Yes D 20.3 0.6 Heptane 2.0 Yes D 23.6 0.6 Toluene 1.0 Yes D 21.3 0.6 CH₂Cl₂ 1.0 Yes D 20.7 0.6 Dioxane 6.0 Yes F *Solids were dissolved at 80° C.

XRPD patterns from binary crystallizations with acetone as primary solvent employing a fast cooling procedure afforded four forms A, C, D, and F. Mixtures of acetone with MTBE, THF, and cyclohexane provided Form A material; Form C was obtained from the mixtures of acetone with EtOAc, heptane, toluene, and CH2Cl2. The mixture of acetone with IPAc afforded Form D, and the mixture of acetone with dioxane afforded Form F solids.

XRPD patterns from binary crystallizations with isoamyl alcohol as the primary solvent employing a fast cooling procedure afforded five forms: C, D, E, F, and G. Form C was obtained from crystallizations using MTBE and EtOAc as cosolvents; Form D was obtained from mixtures of isoamyl alcohol with heptane, toluene, and CH2Cl2; Form E was crystallized out of i-AmOH/THF and i-AmOH/cyclohexane; Form G was obtained from i-AmOH/IPAc and Form F was obtained from i-AmOH/dioxane. Form D was the most common form observed from the crystallizations, and Form F was observed only when dioxane was used as an antisolvent.

Binary solvent recrystallizations of the potassium salt with the slow-cooling procedure were performed using three primary solvents (MeCN, acetone, and i-AmOH) and eight cosolvents (MTBE, EtOAc, IPAc, c-hexane, heptane, toluene, CH2Cl2, and dioxane). All solids were analyzed by XRPD to determine the physical form. Two patterns were observed to be Forms C and D respectively with additional peaks present. Other crystallizations provided Forms D, C, or F. Form D was obtained from the following solvent mixtures: MeCN/MTBE, MeCN/IPAc, MeCN/toluene, MeCN/CH2Cl2 and also from the mixtures of i-AmOH with MTBE, IPAc, cyclohexane, heptane, and toluene. Form C was obtained from MeCN/heptane, i-AmOH/EtOAc, and mixtures of acetone with MTBE, EtOAc, IPAc, cyclohexane, heptane, toluene, and CH₂Cl₂. Form F was crystallized from the mixtures of MeCN/dioxane and i-AmOH/dioxane. The solvent mixture of MeCN/EtOH provided amorphous material. Elemental analysis results indicate that the forms observed may not be pure and/or have bound or residual solvents present. Forms C, D, and F were found to be the most common forms of the potassium salt isolated and based on the results, these forms were chosen for scale-up and further characterization. Differences were found in the XRPD patterns and FTIR spectra of the scale-up lots which were attributed to differences in impurity profiles, crystallinity, and form purity.

6.2.11.4. Characterization of Polymorphic Form C of Diazoxide Potassium Salt.

Form C of diazoxide potassium salt is an acetone solvate with a 2:1 ratio of diazoxide/solvent. It is a crystalline form of diazoxide potassium, with endothermic events at 187 and 360° C. in the DSC. The XRPD pattern for Form C is unique compared to all other forms observed. FTIR (ATR) spectroscopy showed differences between forms. 1H NMR spectra were found to be consistent with the structure of diazoxide with a half-molar equivalent of acetone present. NMR data also indicated that the magnetic environment of the diazoxide structure had changed evidenced by a movement in chemical shift of the aromatic and methyl proton resonances. In addition, the resonance due to the amine proton was not observed which suggested deprotonation in solution. Weight loss by TGA was 8.9%, consistent with a half-molar equivalent of acetone, and occurred near 180° C. consistent with the endotherm observed in the DSC experiment. Moisture-sorption analysis conducted at 25° C. from 0 to 90% RH (adsorption) and 85 to 0% RH (desorption) showed Form C to be a hygroscopic solid, showing approximately 47 wt % water at 90% RH which indicated the sample deliquesced. In comparison, Forms D and F (hemihydrate and dioxane solvate) showed approximately 26 and 22 wt % water at 90% RH respectively. XRPD analysis following the moisture-sorption experiment afforded a pattern consistent with Form D. Slurry experiments were performed on 50/50 mixtures of Forms C, D, and F to determine propensity for conversion and to see if a unique form could be generated. Upon slurrying Form C mixtures in ethyl acetate, acetonitrile, and isopropanol conversion to Form D was observed in all solvents. Without wishing to be bound by theory, these results as well as conversion of Form F to Form D in these conditions suggest that Form D is more thermodynamically stable than Form C and F according to Ostwald's Rule of Stages. A thermal-stability experiment on Form C at 60° C. found the form to be stable. Conversion to another form was not observed.

6.2.11.5. Characterization of Polymorphic Form D of Diazoxide Potassium Salt.

Form D of diazoxide potassium salt is a hemihydrate. It is a crystalline form of diazoxide potassium, with endothermic events at 130, 191, and 352° C. in the DSC. FTIR (ATR) spectroscopy showed differences between forms. 1H NMR spectra were found to be consistent with the structure of diazoxide. NMR data also indicated that the magnetic environment of the diazoxide structure had changed evidenced by a movement in chemical shift of the aromatic and methyl proton resonances. In addition, the resonance due to the amine proton was not observed which suggested deprotonation in solution. Weight loss by TGA was 4.5%, consistent with a half-molar equivalent of water, and occurred near 110° C. consistent with the endotherm observed in the DSC experiment. Moisture-sorption analysis conducted at 25° C. from 0 to 90% RH (adsorption) and 85 to 0% RH (desorption) showed Form D to be a hygroscopic, showing approximately 26 wt % water at 90% RH. In comparison, Forms C and F (acetone and dioxane solvates) showed approximately 47 and 22 wt % water at 90% RH, respectively. XRPD analysis following the moisture-sorption experiment afforded a pattern consistent with Form D. Solubility studies were conducted at pH 2, 7, and 12 for Form D and showed 29, 33, and 59 mg/mL respectively. Solubility concentrations were determined using area-percent calculations with an HPLC calibration curve. Slurry experiments were performed on 50/50 mixtures of Forms C, D, and F to determine their propensity for conversion and to see if a unique form could be generated. Upon slurrying mixtures of Form D with Form C or Form F in ethyl acetate, acetonitrile, and isopropanol conversion to Form D was observed in all solvents.

6.2.11.6. Characterization of Polymorphic Form F of Diazoxide Potassium Salt.

Form F of diazoxide potassium salt is a dioxane solvate with a 2:1 ratio of diazoxide/solvent. It is a crystalline form of diazoxide potassium, with endothermic events at 191 and 363° C. in the DSC. The XRPD pattern for Form F is unique compared to all other forms observed. FTIR (ATR) spectroscopy showed differences between forms. ¹H NMR spectra were found to be consistent with the structure of diazoxide with a half-molar equivalent of dioxane present. NMR data also indicated that the magnetic environment of the diazoxide structure had changed as evidenced by a movement in chemical shift of the aromatic and methyl proton resonances. In addition, the resonance due to the amine proton was not observed which suggested deprotonation in solution. Weight loss by TGA was 13.1%, consistent with a half-molar equivalent of dioxane, and occurs near 180° C. consistent with the endotherm observed in the DSC experiment. Moisture-sorption analysis conducted at 25° C. from 0 to 90% RH (adsorption) and 85 to 0% RH (desorption) showed Form F to be a hygroscopic solid, showing approximately 22 wt % water at 90% RH. In comparison, Forms C and D (acetone solvate and hemihydrate) showed approximately 47 and 26 wt % water at 90% RH, respectively. XRPD analysis following the moisture-sorption experiment afforded a pattern consistent with Form D. Slurry experiments were performed on 50/50 mixtures of Forms C, D, and F to determine their propensity for conversion and to see if a unique form could be generated. Upon slurrying mixtures of Form F with Form C and Form D in ethyl acetate, acetonitrile, and isopropanol conversion to Form D was observed in all solvents.

B. In vivo Obesity Testing 1. Obesity Animal Model

Formulations of salts of any of the compounds of Formulae I-IV prepared as described herein can be tested for efficacy in an animal model of obesity as described by Surwit et al. (Endocrinology 141:3630-3637 (2000)). Briefly, 4-week-old B6 male mice are housed 5/cage in a temperature-controlled (22° C.) room with a 12-h light, 12-h dark cycle. The high fat (HF) and low fat (LF) experimental diets contain 58% and 11% of calories from fat, respectively. A group of mice are fed the HF diet for the first 4 weeks of the study; the remaining 15 mice are fed the LF diet. The mice assigned to the LF diet are maintained on this diet throughout the study as a reference group of lean control mice. At week 4, all HF-fed mice a reassigned to 2 groups of mice. The first group remains on the HF diet throughout the study as the obese control group. The remaining 3 groups of mice are fed the HF diet and administered the controlled release formulation of salts of any of the compounds of Formulae I-IV at about 150 mg of active per kg per day as a single dose administered by oral gavage. Animals are weighed weekly, and food consumption is measured per cage twice weekly until the diets are changed at week 4, whereupon body weight and food intake are determined daily. The feed efficiency (grams of body weight gained per Cal consumed) is calculated on a per cage basis. Samples for analysis of insulin, glucose, and leptin are collected on day 24 (4 days before the diets are changed), on day 32 (4 days after the change), and biweekly thereafter. In all cases food is removed 8 h before samples are collected. Glucose is analyzed by the glucose oxidase method. Insulin and leptin concentrations are determined by double antibody RIA. The insulin assay is based on a rat standard, and the leptin assay uses a mouse standard. At the termination of the study, a postprandial plasma sample is collected and analyzed for triglyceride and nonesterified fatty acid concentrations. After 4 weeks of drug treatment, a subset of 10 animals from each group is killed. The epididymal white adipose tissue (EWAT), retroperitoneal (RP) fat, interscapular brown adipose tissue (IBAT) fat pads, and gastrocnemius muscle are removed, trimmed, and weighed. The percent body fat is estimated from the weight of the epididymal fat pad. A subset of five animals from each group is injected i.p. with 0.5 g/kg glucose. At 30 min post injection, a plasma sample is collected and analyzed for glucose content by the glucose oxidase method.

2. Treatment of Obesity in Humans

Formulations of salts of any of the compounds of Formulae I-IV prepared as described herein can be tested for efficacy in obese humans, as described by Alemzadeh (Alemzadeh et al., J Clin Endocr Metab 83:1911-1915 (1998)). Subjects consist of moderate-to-morbidly obese adults with a body mass index (BMI) greater than or equal to 30 kg/m². Each subject undergoes a complete physical examination at the initial evaluation, body weight being measured on a standard electronic scale and body composition being measured by DEXA.

Before initiation of the study, all subjects are placed on a hypocaloric diet for a lead-in period of 1 week. This is designed to exclude subjects who are unlikely to be compliant and to ensure stable body weight before treatment. Up to 50 subjects are tested at each dosage of drug. Daily dosage is set at 100, 200, and 300 mg/day. The daily dose is divided into 2 doses for administration. The dose is administered as either one, two or three 50 mg capsules or tablets at each time of administration. Subjects are dosed daily for up to 12 months. Subjects are reviewed weekly, weighed, and asked about any side effects or concurrent illnesses.

Twenty-four-hour dietary recall is obtained from each subject. The dietary recalls are analyzed using a standard computer software program. All subjects are placed on a hypocaloric diet and encouraged to participate in regular exercise.

Before commencing, and after completion of the study, the following laboratory tests are obtained: blood pressure, fasting plasma glucose, insulin, cholesterol, triglycerides, free fatty acids (FFA), and glycohemoglobin, and measures of rate of appearance and oxidation of plasma derived fatty acids. Additionally, routine chemistry profiles and fasting plasma glucose are obtained weekly to identify those subjects with evidence of glucose intolerance and/or electrolyte abnormalities. Glucose is analyzed in plasma, by the glucose oxidase method.

Insulin concentration is determined by RIA using a double-antibody kit. Cholesterol and triglycerides concentrations are measured by an enzymatic method. Plasma FFA is determined by an enzymatic colorimetric method. SI was assessed by an iv glucose tolerance test (IVGTT) using the modified minimal model. After an overnight fast, a glucose bolus (300 mg/kg) was administered iv, followed (20 min later) by a bolus of insulin. Blood for determination of glucose and insulin is obtained from a contra lateral vein at −30, −15, 0, 2, 3, 4, 5, 6, 8, 10, 19, 22, 25, 30, 40, 50, 70, 100, 140, and 180 min. SI and glucose effectiveness (SG) are calculated using Bergman's modified minimal-model computer program before and after the completion of the study. Acute insulin response to glucose is determined over the first 19 min of the IVGTT, and the glucose disappearance rate (Kg) is determined from 8-19 min of the IVGTT. Body composition is measured by bioelectrical impedance before and at the completion of the study. Resting energy expenditure (REE) is measured by indirect calorimetry after an overnight 12-h fast, with subjects lying supine for a period of 30 min. Urine is collected over the corresponding 24 h, for measurement of total nitrogen and determination of substrate use, before and after the study.

3. Treatment of Obesity in Humans by Coadministering Diazoxide and Phentermine

Evaluation of a prolonged co-administration of solid oral dosage form of salts of any of the compounds of Formulae I-IV thereof in combination with phentermine can be conducted in humans with moderate-to-morbid obesity and a body mass index (BMI) greater than or equal to 30 kg/m2. Each subject undergoes a complete physical examination at the initial evaluation, body weight being measured on a standard electronic scale and body composition by DEXA.

Before initiation of the study, all subjects are placed on a hypocaloric diet for a lead-in period of 1 week. This is designed to exclude subjects who are unlikely to be compliant and to ensure stable body weight before treatment. Up to 100 subjects are tested. Daily dosage of salts of any of the compounds of Formulae I-IV is set at 200 mg. The daily dose is divided into 2 doses for administration. The dose is administered as either a 100 mg capsule or a 100 mg tablet at each time of administration. Subjects are dosed daily for up to 12 months. Phentermine is administered as a single daily dose of 15 mg. Subjects are reviewed every two weeks, weighed, and asked about any side effects or concurrent illnesses.

All subjects are continued on a hypocaloric diet and encouraged to participate in regular exercise. Before commencing, and after completion of the study, laboratory tests as described in the example above are obtained.

4. Prevention of Diabetes in Prediabetic Humans

The example describes use of salts of any of the compounds of Formulae I-IV in a prediabetic subject to prevent the occurrence of diabetes. Subjects included in the study all have elevated risk of developing diabetes as measured by one of two methods. In a fasting glucose assay they have plasma glucose values between 100 and 125 mg/dl indicating impaired fasting glucose, or in an oral glucose tolerance test they have plasma glucose values between 140 and 199 mg/dl at 2 hours post-glucose load indicating they have impaired glucose tolerance. Treatment is initiated in any subject meeting either criteria. Treated subjects receive either 200 mg diazoxide per day as a 100 mg capsule or tablet twice per day or as two 100 mg capsules or tablets once per day. Placebo treated subjects receive either one placebo capsule or tablet twice per day or two placebo capsules or tablets once per day.

Treatment is continued for once year with OGTT or fasting glucose measured monthly.

5. A sustained Release Coformulation of diazoxide HCl and metformin HCl use to Treat Diabetic Patients

A sustained release co-formulation of diazoxide HCl and metformin HCl is produced by forming a compressed tablet matrix that includes 750 mg of metformin HCl and 100 mg of diazoxide HCl. These active ingredients are blended with sodium carboxymethyl cellulose (about 5% (w/w)), hypromellose (about 25% (w/w), and magnesium stearate (<2% (w/w)). The compressed tablet is further coated with a combination of ethylcellulose (80% (w/w)) and methyl cellulose (20% (w/w)) as a thin film to control rate of hydration and drug release.

Type II diabetic patients are treated with the oral dosage form by administration of two tablets once per day or one tablet every 12 hours. Treatment of the patient with the drug is continued until one of two therapeutic endpoints is reached, or for so long as the patient derives therapeutic benefit from administration. The two therapeutic endpoints that would serve as the basis for the decision to cease treatment include the patient reaching a Body Mass Index (BMI (kg/m2)) between 18 and 25 or the re-establishment of normal glucose tolerance in the absence of treatment. The patient is monitored periodically for (a) glucose tolerance using an oral glucose tolerance test, (b) glycemic control using a standard blood glucose assay, (c) weight gain or loss, (d) progression of diabetic complications, and (e) adverse effects associated with the use of these active ingredients.

6. Prevention or Treatment of Weight Gain in a Patient Treated with Olanzapine P Pharmacotherapy for schizophrenia is initiated for a patient meeting DSM M-III-R criteria for schizophrenia. The patient is administered 10 mg of olanzapine (Zyprexa, Lilly) once per day. Adjunctive therapy to the patient for schizophrenia includes 250 mg equivalent of valproic acid as divalproex sodium (Depakote, Abbott Labs). Weight gain, dyslipidemia and impaired glucose tolerance, and metabolic syndrome are high frequency adverse events in patients treated with this combination of anti-psychotics. Weight gain, dyslipidemia, impaired glucose tolerance or metabolic syndrome are treated by the co-administration of a therapeutically effective dose of a KATP channel opener. The patient is treated with administration of 200 mg/day of salts of any of the compounds of Formulae I-IV as a once daily tablet formulation. Administration of salts of any of the compounds of Formulae I-IV continues until the weight gain, dyslipidemia, impaired glucose tolerance or metabolic syndrome is corrected or until treatment of the patient with olanzapine is discontinued. Dyslipidemia is detected by measuring circulating concentrations of total, HDL, and LDL cholesterol, triglycerides and non-esterified fatty acids. Impaired glucose tolerance is detected through the use of oral or IV glucose tolerance tests. Metabolic syndrome is detected by measuring its key risk factors including central obesity, dyslipidemia, impaired glucose tolerance, and circulating concentrations of key proinflammatory cytokines.

7. Comparison of Single Doses of Diazoxide Administered as Proglycem® Oral Suspension or as Diazoxide Choline Controlled-Release Tablets in Obese Subjects.

7.1 Experimental Design

7.1.1. Objective of Study.

Clinical studies employing a randomized, open-label, parallel protocol comparing the safety, tolerability and bioavailability (i.e., pharmacokinetics) in obese subjects of Proglycem® (oral suspension) and diazoxide choline salt (controlled-release tablet) were conducted. The study evaluated the safety and tolerability of a single 200 mg does (approximately 2 mg/kg) of diazoxide. The study further compared the single dose pharmacokinetics of an oral suspension of the free base of diazoxide (Proglycem®) with a controlled-release tableted formulation of diazoxide choline salt under fasting conditions in obese subjects.

7.1.2. Rationale for Study.

Diazoxide choline was selected as an alternative molecule for the oral administration of diazoxide because of significantly greater aqueous solubility over diazoxide free base, rapid conversion to diazoxide base on exposure to an aqueous environment, and incompatibility with incorporation into sustained release formulations. The fate of diazoxide choline in an aqueous medium in vitro has been extensively characterized. Without wishing to be bound by theory, once solubilized from the controlled-release tablet formulation, prior to absorption, the salt hydrolyzes to the free base of diazoxide and choline hydroxide. This was extensively characterized using UV/Vis absorbance, and occurred at all physiological relevant pH values from pH 2.0 to pH 8.5. This hydrolysis occurred in deionized water and buffered aqueous solutions, and in polar solvents that contained trace amounts of water. Thus, diazoxide, as the free base, is the molecular form absorbed following oral administration of the choline salt of diazoxide to animals or humans. Serum and plasma assays used in TK and PK analysis measure diazoxide free base concentrations. Dissolution assays measure diazoxide free base. Differences in plasma concentration-time profiles of diazoxide will be dependent on the time course of release of diazoxide choline from the tablet matrix in the intestinal tract. Historically, both oral suspension and immediate release capsule formulations of diazoxide have been commercially available, marketed as Proglycem®. The oral suspension has been shown to be highly bioavailable and rapidly absorbed upon administration. Because of the difficulty in characterizing dissolution from the oral suspension, the in-vitro dissolution of Proglycem® capsules was compared under standardized conditions to that of diazoxide choline controlled-release tablets (50 mg and 200 mg).

7.1.3. Inclusion Criteria.

Inclusion criteria for the present study included age 18 to 65 years old inclusive and BMI between 30 and 45 kg/m2, inclusive, and signed informed consent. Female participants were required to be either postmenopausal for at least 1 year, surgically sterile [bilateral tubal ligation, bilateral oophorectomy, or hysterectomy], or practicing a medically acceptable method of birth control. Inclusion criteria further included freedom from serious medical disorders involving the kidneys, digestive system, heart and blood vessels, lungs, liver, eyes, nerves, brain, skin, endocrine system, bones or blood. Subjects, other than their obese condition, were generally healthy as documented by medical history, physical examination, vital sign, 12-lead ECG, and clinical laboratory assessments. Characteristics of the subjects randomized in the study are provided in Table 33.

TABLE 33 Characteristics of subjects randomized in the study Proglycem Oral Diazoxide Choline Controlled- Suspension (n = 15) Release Tablet (n = 15) Parameter Mean ± SD (range) Mean ± SD (range) Age (yr) 32.5 ± 12.1 (18-56) 27.9 ± 12.6 (19-54) BMI 32.9 ± 4.3 (30.2-44.8) 33.8 ± 3.2 (31.4-42.1) (kg/m²) Gender 9/6 7/8 (male/ female)

7.1.4. Exclusion Criteria.

Exclusion criteria included treatment with an investigational drug within 28 days prior to dosing, presence or history of a clinically significant disorder, clinical laboratory test values outside of the accepted reference range, reactive screen for HBV, HCV, or HIV, use of any medication affecting body weight, lipid or glucose metabolism within 2 months, use of any systemic prescription medication from 14 days prior to screening to dosing except hormonal contraceptives, use of any drug known to induce or inhibit hepatic drug metabolism from 28 days prior to screening until dosing, positive test for drug of abuse, current tobacco use, positive pregnancy screen, or pregnant or breastfeeding.

7.1.5. Randomization.

A total of 30 subjects were randomized in the present study. Fifteen subjects were randomized to each arm. For convenience, the 30 subjects were broken up into two cohorts. Cohort 1 checked into the clinic on Oct. 12, 2006, was dosed on Oct. 14, 2006, remained in the clinic for 72 hours following dose administration, returning at 96 and 120 hours after dose administration. Cohort 1 included 10 subjects randomized to receive Proglycem Oral Suspension and 5 subjects randomized to receive a Diazoxide Choline Controlled-Release Tablet. Cohort 2 checked into the clinic on Oct. 14, 2006, were dosed on Oct. 16, 2006, remained in the clinic for 72 hours following dose administration, returning at 96 and 120 hours after dose administration. Cohort 2 included 5 subjects randomized to receive Proglycem® Oral Suspension and 10 subjects randomized to receive a diazoxide choline controlled-release tablet. All subjects completed the study.

7.1.6. Dosing.

Subjects randomized to the Proglycem® Oral Suspension arm received a single 200 mg dose (4 mL) taken with 240 mL of room temperature water. Subjects randomized to the diazoxide choline controlled-release tablet arm received a single tablet containing 290 mg of diazoxide choline, which is equivalent to 200 mg of diazoxide. Doses were administered after an overnight fast, and fasting continued until approximately 4.25 hours after dose administration at which time a standardized meal was served.

7.1.7. Safety Monitoring.

Clinical laboratory tests including hematology, clinical serum chemistry and urinalysis were conducted at screening and at end of study. Hematology included measurements of hemoglobin, hematocrit, white blood cell count with differential, red blood cell count and platelet count. Clinical serum chemistry included evaluation of sodium, potassium, BUN, creatinine, total bilirubin, total protein, albumin, alkaline phosphatase, AST, ALT, glucose, total cholesterol, HDL-cholesterol, and triglycerides. Urinalysis included pH, specific gravity, protein, glucose, ketones, bilirubin, blood, nitrites, urobilinogen, leukocytes, and microscopic urine analysis if sample was dipstick positive. All adverse events were recorded. Vital signs were collected at screening, at one hour prior to dosing, and after dose administration at 1, 3, 6, 9, 12, 24, 48, 72 and 120 hours. Continuous two-lead cardiac monitoring (telemetry) was performed on subjects from 24 hours prior to dose administration (to establish baseline data) until 24 hours after dose administration. 7.1.8. Exploratory Endpoints.

Three exploratory endpoints were evaluated in the study. Blood glucose, insulin and non-esterified fatty acids (NEFA) were measured prior to dosing and at 1, 3, 6, 9, 12, 24, and 48 hours post dosing. Blood glucose and insulin levels were also assessed at end of study. Data collected at times 1, 3, 24, and 48 hours after dose administration were obtained under fasting conditions. The remaining data points were post-prandial measures. Samples for pharmacokinetic analysis to evaluate bioavailability were collected prior to dosing and after dose administration at 1, 2, 4, 6, 8, 12, 16, 20, 24, 32, 40, 48, 72, 96, and 120 hours.

7.2. Results

7.2.1. Adverse Events.

The adverse events observed with each of the two formulations are summarized in Tables 34 and 35. With the single exception, a moderate headache requiring treatment with acetaminophen in a subject treated with Proglycem® Oral Suspension, all adverse events were mild. Both formulations appeared to be well tolerated in the study. One subject administered with diazoxide choline controlled-release tablet experienced mild loss of appetite. Because most obese and obese diabetic animal model studies of diazoxide show some reduction in feed consumption, loss of appetite was, therefore, considered part of the pharmacodynamic response to the drug in obese patients, rather than an adverse event.

TABLE 34 Adverse events (AE) in subjects administered Proglycem ® Oral Suspension (n = 15) Hours Post Corrective Out- Adverse Event Dosing si Treatment come Gastrointestinal Nausea^(a) 2.75 Mild None Resolved Neurological Headache 20.0 Moderate Therapy required Resolved acetaminophen Headache^(b) 6.25 Mild None Resolved Dizziness^(c) 1.0 Mild Assisted to supine Resolved position Dizziness^(c) 3.25 Mild Assisted to supine Resolved position Dizziness^(d) 1.0 Mild Assisted to supine Resolved position Dizziness^(d) 4.25 Mild Assisted to supine Resolved position Miscellaneous Chills^(b) 6.25 Mild None Resolved Dermatitis from 4.75 Mild None Resolved ECG patches^(a) Number of subjects with at least one AE = 5, AEs followed by the same superscript letter are the same subject

TABLE 35 Adverse events (AE) in subjects administered Diazoxide Choline Controlled-Release Tablet (n = 15) Hours Post Corrective Out- Adverse Event Dosing Treatment come Gastrointestinal Nausea^(e) 1.5 Mild None Resolved Dry Heaves^(e) 1.6 Mild None Resolved Neurological Lightheaded 12.0 Mild Assisted to supine Resolved position Dizziness^(g) 20.0 Mild None Resolved Miscellaneous Chills^(f) 2.0 Mild None Resolved Back Pain^(f) 2.0 Mild None Resolved Warm^(g) 20.0 Mild None Resolved Number of subjects with at least one AE = 4, AEs followed by the same superscript letter are the same subject

In addition, a randomized double-blind, placebo-controlled clinical trial was conducted in 24 healthy volunteers. They were randomized in a 2:1 ratio of treated to placebo. Treated subjects received 300 mg/day diazoxide administered as 3 tablets of the controlled release formulation U described in Table 2. The dose was administered once per day in the morning for a period of 7 days.

There were more adverse events reported per subject in the placebo controls than in the diazoxide choline treated arm (Table 35′). The majority of the events were mild. The frequency of the most common adverse events was identical in the diazoxide choline treated arm and in the controls.

TABLE 35 Adverse events (AE) in subjects administered Diazoxide Choline (300 mg Diazoxide) Controlled-Release Tablet or Placebo Adverse Event Diazoxide choline Placebo Headache 4 (25%) 2 (25%) Nausea 3 (19%) 2 (25%) Dizziness 2 (13%) 1 (13%) n 16 8

7.2.2. Clinical Chemistry.

A summary of fasting glucose, fasting insulin, NEFA, and serum sodium (Na), potassium (K) and creatinine level is provided in Table 36. Neither treatment resulted in significant changes in serum Na, K or creatinine from baseline to end of study. Treatment with diazoxide primarily impacts glucose-stimulated insulin secretion rather than basal insulin secretion. Fasting glucose levels increased slightly in the first 3 hours following dose administration. This increase was more pronounced in the Proglycem® Oral Suspension arm as compared to the diazoxide choline controlled-release tablet arm. These results are consistent with the differences in measured rates of dissolution from these formulations (see Table 5 herein) and the PK levels (see FIGS. 24 and 25 herein). In this study, there is no evidence for a reduction in fasting insulin after the administration of the single dose of diazoxide. There is also no evidence for a significant increase in fasting glucose as measured at 24 or 48 hours after dose administration. The most sensitive pharmacodynamic response measure is NEFA. Diazoxide treatment normally results in a short-term increase in NEFA, which returns to levels at or below baseline in about 6 hours. Both formulations showed this transient increase in NEFA. Treatment with Proglycem® Oral Suspension resulted in a statistically significant increase (p<0.001) in NEFA at 3 hours, which by 6 hours after dose administration had returned to levels well below baseline. Similarly the increase in NEFA at 3 hours after administration of diazoxide choline controlled-release tablets was statistically significant (p<0.0012). NEFA levels in the diazoxide choline controlled-release tablet treated subjects also returned to levels well below baseline by 6 hours after dose administration.

TABLE 36 Fasting glucose, fasting insulin, non-esterified fatty acid (NEFA), serum sodium (Na), potassium (K) and creatinine of subjects Diazoxide Choline Proglycem Oral Controlled-Release Suspension (n = 15) Tablet (n = 15) Parameter Mean ± SD (range) Mean ± SD (range) Fasting Glucose (mg/dL) Baseline 93 ± 6 (83-104) 93 ± 6 (83-108) 1 h post-dose 99.4 ± 7 (89-117) 97.9 ± 7 (92-117) 3 h post-dose 102.8 ± 6 (93-113) 98.0 ± 7 (90-112) 24 h post-dose 97 ± 5 (87-106) 98 ± 8 (88-118) 48 h post-dose 92 ± 5 (84-103) 93 ± 5 (83-104) Fasting Insulin (μIU/ml) Baseline 8.6 ± 5.2 (2.9-24.1) 9.9 ± 3.3 (4.4-16.3) 24 h post-dose 12.0 ± 8.9 (6.1-41.9) 11.1 ± 3.3 (5.5-17.0) 48 h post-dose 9.5 ± 5.7 (4.0-28.9) 11.1 ± 2.9 (5.3-17.8) NEFA (μmol/L) 1 h post-dose 0.36 ± 0.11 (0.21-0.65) 0.34 ± 0.14 (0.09-0.67) 3 h post-dose 0.53 ± 0.13 (0.31-0.78) 0.55 ± 0.19 (0.28-0.88) 6 h post-dose 0.13 ± 0.07 (0.07-0.32) 0.11 ± 0.04 (0.07-0.21) 24 h post-dose 0.36 ± 0.10 (0.23-0.62) 0.40 ± 0.18 (0.25-0.97) 48 h post-dose 0.38 ± 0.13 (0.2-0.66) 0.39 ± 0.15 (0.17-0.65) Na (mEq/L) Baseline 138.0 ± 2.0 (135-142) 138.1 ± 2.6 (134-143) End of Study 138.7 ± 1.1 (137-141) 139.3 ± 2.3 (135-144) K (mmol/L) Baseline 4.4 ± 0.4 (3.9-5.5) 4.2 ± 0.3 (3.8-5.1) End of Study 4.4 ± 0.3 (3.9-5.2) 4.2 ± 0.3 (3.8-5.2) Creatinine (mg/dL) Baseline 0.9 ± 0.1 (0.7-1.1) 0.9 ± 0.2 (0.7-1.4) End of Study 1.0 ± 0.1 (0.8-1.3) 0.9 ± 0.2 (0.6-1.3)

All other clinical laboratory tests, including hematology, clinical serum chemistry, and urinalysis were within normal ranges for obese subjects.

7.2.3. Vital Signs.

A graph depicting sitting blood pressure at baseline and at various times following dose administration is provided in FIG. 22. Neither treatment was associated with a sustained reduction or increase in blood pressure, nor was there any clear trend from baseline over 6, 9, 12, or 24 hours after dose administration. Pulse rates dropped incrementally from baseline levels in the period from dose administration to 3 hours post dosing (FIG. 23). Pulse rates rose to levels equivalent to baseline or slightly above baseline in the period from 6 to 12 hours after dose administration, and remained at baseline levels from 24 hours after dose administration to the end of the study (FIG. 23). Review of the baseline cardiac telemetry data for all subjects was conducted the morning of dosing. Based on the absence of clinical significant abnormalities, all subjects were allowed to proceed with dosing. The cardiac telemetry data from study time 0 to 24 hours after dose administration was evaluated. No clinically significant abnormalities (e.g. arrhythmias) were observed.

7.2.4. Preliminary Evaluation of Diazoxide Plasma Pharmacokinetics.

Compared to the Proglycem® Oral Suspension, the diazoxide choline controlled-release tablet had a 30% lower peak exposure (Cmax) and a 15% lower total exposure (AUC) when the formulations were administered orally as 200 mg equivalents of diazoxide (Table 37). The timing of the peak plasma concentration (Tmax) occurred at a median time of 4 hr after administration of the oral suspension and 20 hr after administration of the diazoxide choline controlled-release tablet. The terminal half-life for diazoxide was similar with both formulations (29 hr for the oral suspension and 32 hr for the tablet). The two formulations had similar between-patient variability for Cmax and AUC. The most notable difference between the two formulations following a single dose was the lower, broader peak of the concentration-time profile of the tablet formulation (FIGS. 24 and 25). The peak concentration for the tablet averaged about 30% less and occurred about 16 hours later than the suspension. FIGS. 24 and 25 illustrate that the mean peak concentration was virtually unchanged between 12-hr and 24-hr post-dose following administration of the diazoxide choline controlled-release tablet formulation. Because of a more sustained release pattern, the tablet formulation was predicted to have a greater accumulation factor with chronic dosing than was the oral suspensions (3.96 vs. 2.84). Simulations of repeated once-daily dosing with the two formulations (assuming linear pharmacokinetics) predicted that the two formulations would have similar trough concentrations at steady-state (22-23 μg/mL) (see FIG. 26).

TABLE 37 Summary Statistics for Plasma Diazoxide Pharmacokinetic Parameters after a Single Dose of Two Formulations Cmax Tmax AUC(0-24) AUC(0-120) AUC(0-∞) λz T½ Accum μg/mL hr μg · hr/mL μg · hr/mL μg · hr/mL 1/hr hr Factor Diazoxide Choline Controlled-Release Tablets N 15 15 15 15 15 15 15 15 Mean 9.25 22.1 166 553 618 0.0242 31.9 3.96 Geomean 9.07 19.6 157 531 588 0.0229 30.2 3.74 St Dev 1.89 11.5 54 160 198 0.0085 10.6 1.48 SEM 0.49 3.0 14 41 51 0.0022 2.7 0.38 CV % 20.5% 52.0% 32.5% 28.9% 32.0% 35.2% 33.3% 37.5% Median 9.08 20.0 164 518 586 0.0237 29.3 3.80 Min 5.53 8.00 74.6 261 274 0.0144 15.9 2.26 Max 13.2 48.0 266 842 1024 0.0437 48.3 7.84 Diazoxide Oral Suspension (Proglycem) N 15 15 15 15 15 15 15 15 Mean 13.5 6.67 245 643 696 0.0256 28.6 2.84 Geomean 13.3 5.36 243 629 677 0.0249 27.9 2.79 St Dev 2.3 5.49 35 142 173 0.0068 6.3 0.57 SEM 0.6 1.42 9 37 45 0.0018 1.6 0.15 CV % 17.1% 82.3% 14.4% 22.1% 24.9% 26.7% 22.0% 20.0% Median 13.3 4.00 245 620 671 0.0234 29.6 2.86 Min 10.1 2.00 183 464 488 0.0180 16.1 1.84 Max 18.0 24.0 314 926 1054 0.0430 38.5 4.25

7.3 Conclusions of Comparison of Single Doses of Diazoxide Administered as Proglycem® Oral Suspension or as Diazoxide Choline Controlled-Release Tablets in Obese Subjects.

The present clinical study on the comparison of a single dose of diazoxide choline controlled-release tablet with an equivalent dose of diazoxide administered as Proglycem® Oral Suspension in obese patients indicates that both formulations were well tolerated. With a single exception, moderate headache requiring treatment with acetaminophen in one subject treated with Proglycem®, all adverse events were mild. Diazoxide choline controlled-release tablets appear to have a better CNS safety profile than Proglycem® Oral Suspension as evidenced by the absence of headaches and reduced rate of dizziness. Exploratory endpoints showed no detrimental impact. Preliminary pharmacokinetic analysis showed that compared to Proglycem® Oral Suspension, the diazoxide choline controlled-release tablet had a 30% lower peak exposure (Cmax) and a 15% lower total exposure (AUC) for the same 200 mg diazoxide equivalent dose. The timing of the peak plasma concentration (Tmax) occurred at a median time of 4 hr after administration of the oral suspension and 20 hr after administration of the diazoxide choline tablet. The terminal half-life for diazoxide is similar with both formulations (29 hr for the oral suspension and 32 hr for the tablet). The two formulations had similar between-patient variability for Cmax and AUC.

8. Increasing Insulin Sensitivity

Type II diabetes is characterized by increasing insulin resistance, ultimately followed in the progression of the disease by beta cell exhaustion and insulin dependence. Improvements in insulin sensitivity without contributing to beta-cell exhaustion are critical to management of type II diabetes. Likewise, preservation of beta-cell mass in newly diagnosed type I diabetics by KATP channel openers is dependent on the ability to provide beta-cell rest. In non-insulin dependent individuals, direct evidence of beta-cell rest is provided by reductions in fasting insulin. In insulin dependent individuals, insulin secretion is measured indirectly as basal c-peptide levels.

A controlled clinical study was conducted to evaluate the potential for Diazoxide Choline Controlled-Release Tablets administered once daily to improve insulin sensitivity and provide beta-cell rest as evidenced by reductions in homeostasis model assessment-insulin resistance (HOMA-IR) and reductions in fasting insulin. Up to 8 obese individuals with or without hypertension or dyslipidemia, who may also have impaired fasting glucose or impaired glucose tolerance were randomized to received no drug, or diazoxide equivalent doses of 100 mg/day, 200 mg/day or 300 mg/day for 14 days administered as Diazoxide Choline Controlled-Release Tablets. All subjects were placed on a reduced calorie diet. Fasting insulin was measured pre-dose and on day 7, when diazoxide was expected to have achieved steady state pharmacokinetics. Insulin resistance was reduced in a dose dependent manner by day 7 (Table 38). Reductions in fasting insulin were realized at all dose levels (Table 39). Thus, treatment with Diazoxide Choline Controlled-Release Tablets once per day result is rapid and dramatic improvements in insulin resistance which combined with the beta cell rest provided by reductions in fasting insulin result in preservation of beta cell mass and function while improving glycemic control.

TABLE 38 Changes in HOMA-IR in treated obese subjects Homa-IR Per- Pre-dose Day 7 centage Cohort N Mean SEM Mean SEM Change Control 6 2.3 0.51 2.7 0.47 17% 100 mg/day 7 4.0 0.73 2.8 0.52 −30% 200 mg/day 5 3.1 0.80 2.1 0.66 −32% 300 mg/day 8 3.2 0.49 2.0 0.44 −38%

TABLE 39 Changes in fasting insulin in treated obese subjects Fasting Insulin ( μIU/mL) Pre-dose Day 7 Percentage Cohort N Mean SEM Mean SEM Change Control 6 10.9 2.41 13.1 2.13   20% 100 mg/day 7 18.2 3.31 11.7 1.97 −36% 200 mg/day 5 13.7 3.45  9.1 2.72 −34% 300 mg/day 8 13.4 1.97  7.8 1.61 −42%

9. Relationship Between PK Parameters and Weight and Body Mass Index (BMI)

The historic data relating PK parameters to weight and BMI for the Proglycem oral suspension product suggested that there was a strong negative relationship between diazoxide peak drug levels (Cmax) and diazoxide area under the curve (AUC) for single doses and the weight and body mass index of dosed patients. A strong negative relationship between these key PK parameters would result in the need to adjust dose by patient in order to achieve consistent pharmacokinetic profiles in treated patients.

The relationship between Cmax and AUC and the weight and BMI of subjects receiving either Proglycem or Diazoxide Choline Controlled-Release Tablets was evaluated in a controlled clinical study. Fifteen overweight or obese subjects with weight ranging from 172 lbs to 345 lbs and BMI ranging from 30.2 to 44.8 were randomized to receive a single 200 mg dose of diazoxide administered as Proglycem oral suspension. Fifteen overweight or obese subjects with weight ranging from 173 lbs to 288 lbs and BMI ranging from 31.4 to 42.1 were randomized to receive a single 200 mg dose of diazoxide administered as a Diazoxide Choline Controlled-Release Tablet. Circulating diazoxide levels were measured on samples collected prior to dosing and after dose administration at 1, 2, 4, 6, 8, 12, 16, 20, 24, 32, 40, 48, 72, 96, and 120 hours.

The correlation between weight and Cmax and AUC and between BMI and Cmax and AUC were evaluated within each of the dosed cohorts of subjects. There was a significant negative relationship between weight and AUC, weight and Cmax and between BMI and Cmax in the Proglycem treated subjects (Table 40). No such relationship existed between either weight or BMI and AUC or Cmax in the Diazoxide Choline Controlled-Release Tablet treated subjects (Table 41).

TABLE 40 Relationship between PK Parameters and Weight/BMI for Proglycem Proglycem Oral Suspension AUC Cmax Wt −0.56* −0.75** BMI −0.31 −0.61* (*p < 0.05, **p < 0.01)

TABLE 41 Relationship between PK Parameters and Weight/BMI for Proglycem Diazoxide Choline Controlled-Release Tablet AUC Cmax Wt −0.28 −0.23 BMI −0.04 0.17

10. Clinical Studies of Diazoxide Treatment of Dyslipidemia

Seven obese subjects including those with and without dyslipidemia were dosed for 7 days with the equivalent of 400 mg per day of diazoxide administered as diazoxide choline controlled-release tablets QD (two tablets taken daily, each containing 200 mg equivalent diazoxide per tablet prepared as formula K′ (Table 2)). Subsequently, they were withdrawn from treatment and monitored for an additional 7 days.

The results of lipid profile taken on day 7 of treatment and on day 7 of monitoring (post treatment) are shown as an average for the treated individuals in Table 42. On day 7 of treatment, treated individuals averaged 8.6% reduction in total cholesterol, an 8.7% increase in HDL cholesterol and a 57.6% reduction in circulating triglycerides. At the end of the 7 day monitoring period, there was a reversal of each of these impacts; Total cholesterol was only 6.9% below baseline, HDL cholesterol had returned to 1 mg/dL below baseline and circulating triglycerides were now 44.4% below baseline. The most dramatic changes were realized in a 46 year old male who started treatment with total cholesterol of 342 mg/dL, HDL cholesterol of 37 mg/dL and circulating triglycerides of 1221 mmol/L. Following treatment, this subject realized a reduction in total cholesterol of 60 mg/dL, increase in HDL cholesterol of 15 mg/dL and a reduction in circulating triglycerides of 980 mmol/L. Seven days after discontinuation of treatment, this individual's total cholesterol had risen by 11 mg/dL, HDL cholesterol had dropped by 4 mg/dL and circulating triglycerides had risen by 124 mmol/L mg/dL.

TABLE 42 Changes in blood lipid parameters in a group of treated obese dyslipidemic patients Day 7 of 7 Days post- Variable Baseline treatment dosing Total Cholesterol 233 213 217 (mg/dL) HDL Cholesterol 46 50 45 (mg/dL) Triglycerides (mmol/L) 304 129 169

11. Clinical Studies of Diazoxide Treatment of Hypertriglyceridemia

Individual patients with elevated fasting triglycerides, with or without elevated total cholesterol or LDL-cholesterol, were treated with diazoxide, administered as Diazoxide Choline Controlled-Release Tablets for 7 to 49 days. All patients were dosed once per day for an uninterrupted period of time. The dosages were achieved by combinations of two tablets, a 50 mg diazoxide equivalent dose tablet of diazoxide choline made as formula J (Table 2) and a 200 mg diazoxide equivalent dose tablet of diazoxide choline made as formula K′ (Table 2). Table 43 provides this information.

TABLE 43 Combinations of Diazoxide Choline Controlled-Release Tablets Diazoxide equivalent dose Tablets of J Tablets of K′ 100 mg 2 0 150 mg 3 0 200 mg 0 1 250 mg 1 1 300 mg 2 1 350 mg 3 1 400 mg 0 2

Several patients (subjects 1-6 and 8-9) were dosed for day 7 or 14 days at the diazoxide equivalent doses indicated in Table 44. Patient 7 received a diazoxide equivalent dose of 150 mg for 5 days followed by 9 days at a diazoxide equivalent dose of 300 mg (Table 44). Several patients, subjects 10, 12 and 13, were treated for 7 days at a diazoxide equivalent dose of 150 mg, followed by 7 days at a diazoxide equivalent dose of 200 mg, followed by 7 days at a diazoxide equivalent dose of 250 mg, followed by 7 days at a diazoxide equivalent dose of 300 mg, followed by 21 days at a diazoxide equivalent dose of 350 mg (Table 45). One patient, number 11, was treated by titrating the drug using diazoxide equivalent doses of 150 mg, 200 mg, 250 mg and 300 mg for 7, 14, 7 and 17 days, respectively. Patients were placed on a low fat diet at the start of treatment which continued to the end of treatment. A control subject group which did not receive diazoxide met the elevated triglyceride level of the patient group (above 150 mg/d1 measured in blood under fasting conditions) and were placed on the same low fat diet as the treatment group (Table 46).

Each subject's baseline circulating triglyceride level was calculated as the average of two measurements of fasting triglycerides taken approximately two weeks apart, with the second taken predose on the first day of treatment. Patients treated for 14 days or less had their fasting triglyceride levels measured on days 1, 3, 7, and 14 following the start of dosing. Patients treated for greater than 14 days had their fasting triglyceride levels measured on days 7, 14, 21, 28, 35, 42 and 49 following the start of dosing.

The controls, with a single exception with borderline high triglycerides, on average had higher fasting triglycerides at the end of the diet period than when they started.

TABLE 44 Fasting Triglyceride (mg/dl) Responses in Patients Treated for 7 or 14 days Study Day Percent Subject Dose Baseline 1 3 7 14 change Baseline triglyceride between 150 and 200 mg/dl 2 100 mg 173 139 124 85 109 −37% 3 100 mg 180 231 254 133 181 1% 1 200 mg 168 152 112 129 135 −20% 21 150 mg/ 175 207 109 131 — −25% 300 mg 4 300 mg 181 159 124 113 106 −41% 22 300 mg 183 177 94 — — −48% 6 300 mg 187 188 162 188 157 −16% 5 400 mg 186 245 200 179 — −4% Baseline triglyceride between 200 and 500 mg/dl 7 150 mg/ 205.5 207 145 103 104 −49% 300 mg 23 300 mg 265 152 101 — — −62% 24 300 mg 279 328 217 — — −22% 8 400 mg 273 284 207 174 — −36% Baseline triglyceride above 500 mg/dl 9 400 mg 723 511 279 241 — −67%

TABLE 45 Fasting Triglyceride (mg/dl) Responses in Patients Treated for 35 to 49 days Percent Subject Baseline 7 14 21 28 35 42 49 change Subjects with baseline triglycerides between 150 and 200 mg/dl MSC 153 114 116 111 129 131 132 95 −38% 10 161 169 111 90 108 104 108 99 −39% Subjects with baseline triglycerides between 200 and 500 mg/dl 11 202 181 143 155 137 126 — — −37% 12 222 194 140 169 161 149 134 139 −37% 13 336 184 230 173 172 176 130 127 −62%

TABLE 46 Fasting Triglyceride (mg/dl) Responses in Patients on Low Fat Diet for 7 to 21 Days Study Day Percent Subject Baseline 1 3 7 14 21 change Baseline triglyceride between 150 and 200 mg/dl 14 158 151 131 93 133 113 −28% 15 199 190 292 184 165 203 2% Baseline triglyceride between 200 and 500 mg/dl 16 212 246 194 274 255 246 16% 17 218 223 272 343 251 252 16% 18 234 227 161 229 208 287 23% 19 248 216 202 363 347 342 38% 20 299 425 308 502 275 — −8%

Patients treated with diazoxide equivalent doses of 100 mg per day experienced reductions in fasting triglycerides of 38% at 7 days and 18% at 14 days of treatment. Patients treated with diazoxide equivalent doses of 200 mg per day experienced reductions in fasting triglycerides of 23% at 7 days and 20% at 14 days of treatment. Patients treated with diazoxide equivalent doses of 300 mg per day experienced reductions in fasting triglycerides of 18% at 7 days and 29% at 14 days of treatment. Patients treated with diazoxide equivalent doses of 400 mg per day experienced reductions in fasting triglycerides of 50% at 7 days of treatment. Patients treated with diazoxide equivalent doses of 150 mg and 300 mg for 5 and 9 days, respectively experienced reductions in fasting triglycerides of 50% at 7 and 14 days of treatment. In patients titrated from 150 mg to 300 or 350 mg, fasting triglycerides were reduced by 21%, 31%, 38%, 38% and 49% at the end of treatment with diazoxide equivalent doses of 150 mg, 200 mg, 250 mg, 300 mg and 350 mg, respectively. The response to the drug tended to be more pronounced at any dose level at elevated baseline triglyceride levels. The most pronounced impact on fasting triglycerides were realized in subjects with baseline fasting triglyceride levels greater than 300 mg/dl. There was no consistency tendency for elevations in LDL-cholesterol in treated patients, even those starting with very high fasting triglyceride levels at baseline. For example, patient 8 and 9 had reductions in LDL-cholesterol of −8.6% and −15.3%, respectively. Patient 13, in contrast had a rise in LDL-cholesterol of 6.7% on day 21 and of 11% by day 42.

12. Randomized Double-Blind, Placebo-Controlled Diazoxide Clinical Trial in Healthy Volunteers

A randomized double-blind, placebo-controlled clinical trial was conducted in 24 healthy volunteers. They were randomized in a 2:1 ratio of treated to placebo. Treated subjects received 300 mg/day diazoxide administered as 3 tablets of the formulation U as described in Table 2. The dose was administered once per day in the morning for a period of 7 days. A number of lipid parameters were measured at baseline and again on day 7 including fasting triglycerides, total cholesterol, LDL cholesterol, HDL cholesterol, and non-HDL cholesterol. Additionally, blood pressure was measured on a daily basis.

The lipid response to 7 days treatment with diazoxide choline is provided in table 47. Treatment of subjects with diazoxide choline for 7 days resulted in a reduction of triglycerides of 13.2%. In contrast, in the placebo treated arm there was an increase of 11.7%. There was a reduction in total cholesterol, LDL cholesterol and non-HDL cholesterol from baseline to day 7 in both arms. The reduction in the diazoxide choline treated arm was greater for each lipid parameter than the corresponding reduction in the placebo arm. There was no change in HDL cholesterol in the placebo arm, whereas there was an increase HDL cholesterol of 8.2% in the diazoxide choline treated arm. Seven days of diazoxide choline treatment was associated with a marked decrease in triglycerides, reductions in LDL, non-HDL and total cholesterol and a marked increase in HDL cholesterol in these subjects.

TABLE 47 Changes in lipid parameters associated with 7 days of treatment with diazoxide choline (300 mg diazoxide/day) or placebo. Percent Baseline Day 7 change mean ± mean ± from Treatment Lipid parameter SEM SEM baseline Diazoxide Triglycerides 76 ± 7.8 66 ± 3.9 −13.2% choline (mg/dL) Total cholesterol 175 ± 7.5  171 ± 6.4  −2.3% (mg/dL) LDL cholesterol 111 ± 5.8  105 ± 4.9  −5.4% (mg/dL) HDL cholesterol 49 ± 3.3 53 ± 2.9 8.2% (mg/dL) non-HDL cholesterol 126 ± 6.6  118 ± 5.5  −6.3% (mg/dL) Placebo Triglycerides  60 ± 10.7 67 ± 9.8 11.7% (mg/dL) Total cholesterol 171 ± 13.7 167 ± 9.9  −2.3% (mg/dL) LDL cholesterol 105 ± 11.8 101 ± 11.3 −3.8% (mg/dL) HDL cholesterol 53 ± 4.4 53 ± 3.7 0.0% (mg/dL) non-HDL cholesterol 117 ± 12.1 114 ± 10.6 −2.6% (mg/dL)

There was no change in blood pressure in the placebo treated arm over the 7 days of treatment. In contrast, 7 days of diazoxide choline treatment resulted in a reduction in diastolic blood pressure of 6.2 mmHg. This reduction in diastolic blood pressure has been observed in other studies of diazoxide choline and has persisted through the end of 7 weeks of treatment. The blood pressure response to diazoxide choline treatment in two separate clinical studies is shown in table 48. TR002 utilized 435 mg diazoxide choline achieved from one tablet of formulation K′ (290 mg diazoxide choline) and two tablets of formulation J (each 72.5 mg diazoxide choline). In PK008, the dose was administered as one and one ½ tablet formulation K′.

TABLE 48 Blood pressure changes in response to diazoxide choline treatment in two clinical studies. Diazoxide choline Duration of End of Study dose treatment Parameter Baseline treatment EC004 435 mg  7 days Systolic 113.2 113.5 Diastolic 69.4 63.2 TR002 508 mg 49 days Systolic 118.1 117.0 Diastolic 77.7 67.0

13. Clinical Study in Hypertriglyceridemic Patients

A clinical study was conducted in 12 hypertriglyceridemic patients. Patients received 435 mg/day diazoxide choline (300 mg diazoxide/day equivalent dose), and were dosed once per day in the morning. They were divided into two groups, one initially receiving a formulation which has lower bioavailability (formulation U) and one receiving a formulation with higher bioavailability (formulation K′). After day 8 they were crossed over from one formulation to the other formulation. The dosages were achieved by ingesting 1.5 tablets of formula K′ (200 mg diazoxide equivalent dose tablet of diazoxide choline) and 3 tablets of formula U (100 mg diazoxide equivalent dose tablet of diazoxide choline) (Table 2). Treatment continued for 16 days. The endpoint of the study was percentage change from baseline to day 15 of triglycerides, total cholesterol, LDL cholesterol, HDL cholesterol and non-HDL cholesterol. Lipid parameters were measured at baseline and on days 7 and 15. Blood pressure was collected at multiple time points.

Treatment of hypertriglyceridemic patients with diazoxide choline for 15 days resulted in marked reductions in triglycerides, total cholesterol, LDL cholesterol, and nonHDL cholesterol and a marked increase in HDL cholesterol. The bioavailability of the formulation directly contributed to the therapeutic response, with patients receiving the formulation having higher bioavailability showing improved response to treatment. In general, the longer the patient was treated with diazoxide choline, the greater the therapeutic response. Results are provided in Table 49.

TABLE 49 Lipid responses in diazoxide choline treated hypertriglyceridemic patients. Formulation U Formulation K′ Parameter N Day 7 N Day 15 N Day 7 N Day 15 Triglycerides 6 −22% ± 14% 6 −24% ± 17% 6 −46% ± 10% 6 −39% ± 4% Tot-C 6 −2% ± 2% 6 −9% ± 4% 6 −9% ± 3% 6 −13% ± 6% LDL-C 6  1% ± 4% 6 −11% ± 4%  6 −7% ± 5% 6  −16% ± 11% HDL-C 6  5% ± 13% 6 10% ± 5% 6 12% ± 4% 6  13% ± 4% Non-HDL-C 6 −4% ± 4% 6 −14% ± 4%  6 −15% ± 4%  6 −20% ± 8%

Treatment with diazoxide choline also resulted in lower supine and standing blood pressure. At baseline patients in the study had supine systolic blood pressure of 121 mmHg and supine diastolic blood pressure of 69 mmHg Seven days of treatment reduced supine blood pressure to 116/62.5, an average drop in supine blood pressure of nearly 6 mmHg. Similarly, at baseline patients in the study had standing blood pressure of 123/80 mmHg. After 7 days of treatment, this was reduced to 112/75.5 mmHg, an average drop in standing blood pressure of nearly 8 mmHg.

14. Administration with Food to Reduce Adverse Events

Two studies were conducted to confirm the benefit of administration of diazoxide choline with food to reduce the incidence of adverse events. In each study, the subjects received 435 mg/day of diazoxide choline administered either as one intact 290 mg tablet and two intact 72.5 mg tablets (one tablet of formulation K′ (290 mg diazoxide choline) and two tablets of formulation J) or as one intact 290 mg tablet (formulation K′) and one half of a 290 mg tablet (formulation K′) (latter resulting in a 145 mg dose). The dose was administered either on a fasted basis or within 15 minutes of a meal which included solid food.

In the study where the dose was administered while subjects were fasted, all of the 16 subjects receiving the drug experienced at least one adverse event. In contrast, in the study where the dose was administered with a meal, only 42% of 12 subjects experienced at least one adverse event. In particular, the incidence of gastrointestinal adverse events was substantially reduced including nausea, abdominal pain, abdominal distention, diarrhea, gastrointestinal reflux, and chest pain (table 50). Similarly, the incidence of headaches, edema, and various types of pain were also reduced or prevented when the drug was dosed with solid food. These results were observed despite the fact that there was no food effect on PK. At steady state, there is no difference in average circulating drug levels, Cmax or Tmax when the dose is administered with our without food. Administration with food significantly reduced the incidence of a wide range of adverse events without adversely affecting steady-state pharmacokinetics and therapeutic response. This conclusion is further supported by the data from CS04, in which diazoxide choline was administered with food, where the incidence of adverse events per subject was lower in the drug treated arm than in the placebo treated arm (Table 35).

TABLE 50 Incidence of adverse events when dosed under fasted or fed conditions Incidence under fasted Incidence under fed Adverse Event administration administration Nausea 25% 8% Abdominal pain  6% 0% Abdominal distention  6% 0% Diarrhea  6% 0% GERD  6% 0% Chest pain 18% 0% Headache 68% 8% Pain (extremity, back, neck, 31% 0% joint, etc.) Swelling and edema 38% 17%  15. Study of Diazoxide Choline Titration in Patients with Very High Triglycerides

15.1 Experimental Design

A clinical study was conducted in patients with very high triglycerides to test the effects of diazoxide choline controlled release tablets (DCCR tablets) alone and in combination with fenofibrate choline. The study enrolled 44 male and female ambulatory patients who were at least 18 years old. The primary inclusion criteria included: (1) average fasting triglycerides as measured 7 and 3 days before randomization was >500 mg/dL and <1500 mg/dL; (2) if statin treated at screening, patients must have been willing to switch to Atorvastatin 20 mg at the start of the run-in/washout and continue Atorvastatin 20 mg treatment throughout the study; (3) patients must have been willing to washout of all non-statin lipid lowering drugs; (4) fasting glucose must have been <126 mg/dL and their HbAlc must have been <6.5% at screening. Patients with a history of coronary bypass and/or angioplasty, myocardial infarction or stroke were eligible for enrollment.

Following screening, enrolled patients were subjected to a 6 week run-in/washout period during which subjects discontinued all non-statin lipid lowering medications and became compliant with the NCEP ATPIII TLC diet and lifestyle guidance. Patients not treated with a statin at screening remained statin naïve throughout the study. Patients were then randomized equally to one of two treatment arms: DCCR tablet or placebo. Half of the patients in each treatment arm were treated with atorvastatin (Lipitor®) 20 mg and the other half remained statin naïve.

Patients then entered a titration phase in which they received 145 mg DCCR or its equivalent placebo once daily for 10 days, followed by 217.5 mg DCCR or its equivalent placebo once daily for 10 days, and then 290 mg DCCR or its equivalent placebo for the remainder of the study. Following the titration phase, patients were treated with a maintenance dose for 9 weeks, followed by 6 additional weeks of treatment with the addition of 135 mg fenofibrate choline (Trilpix®) in patients who were statin naïve. The total duration of treatment for all patients was 18 weeks (126 days). Dosing occurred once daily, typically in the morning with food.

Collected patient data included levels of fasting triglycerides, Apo B, total cholesterol, VLDL cholesterol, LDL cholesterol, HDL cholesterol, and non-HDL cholesterol, all measured as percent change from baseline to both day 84 and day 126 (after 12 and 18 weeks of treatment).

DCCR tablets were packaged in Formpack® cold formed blister packs. The blister packs consisted of a cold-formable primary package that included at least one layer of aluminum foil, one layer of polyvinylchloride and one layer of nylon. Blisters were sealed with a flexible aluminum foil affixed with an adhesive, which provided for effective protection of the product from light, moisture, oxygen and other gases. Each tablet of the daily dose was packaged in a separate blister. The blister packaged DCCR was then assembled into a kit that provided clear instruction on dosing and administration. DCCR used in the titration phase of treatment was packaged into a separate titration kit that included the daily dose of each dose strength used during the titration phase. The separate packaging of drug into a titration pack allowed for a high level of patient compliance during the titration phase of the study, thereby improving the likelihood that the patient would not experience an adverse event that might lead to forgoing continued treatment.

15.2 Lipid Changes in DCCR+Fenofibrate Patients vs. Placebo+Fenofibrate Patients

Lipid data collected from patients in the DCCR+fenofibrate arm of the study from baseline to day 126 were compared with lipid data from patients in the placebo+fenofibrate arm from day 84 to day 126, i.e., the period in which patients were treated with placebo and fenofibrate. Patients in the DCCR+fenofibrate arm showed greater median decreases in levels of triglycerides (TG), total cholesterol (TC), non-HDL-C and VLDL-C compared with patients in the placebo+fenofibrate arm. Patients in the DCCR+fenofibrate arm also showed a greater median increase in HDL-C compared with patients in the placebo+fenofibrate arm. However, the DCCR+fenofibrate arm had smaller median increases in levels of LDL-C compared to the placebo+fenofibrate arm. Median percentage changes for each lipid measure is summarized in Table 51 below.

TABLE 51 Median Percentage Changes from Baseline for DCCR vs. Placebo (with addition of fenofibrate) DCCR + Fenofibrate Placebo + Fenofibrate (Median Change) (Median Change) TG −77.8%  −52% TC  −21% −11.3% Non-HDL-C −33.8% −15.9% VLDL-C −84.5% −50.5% LDL-C +41.5% +56.7% HDL-C +37.5% +21.4%

Thus administration of DCCR with fenofibrate choline shows an additive effect for decreasing levels of TG, TC, non-HDL-C and VLDL-C, as well as increasing levels of LDL-C and HDL-C. FIG. 27 shows a graph summarizing the median changes in lipid levels for the DCCR arm compared with placebo arm.

15.3 Lipid Changes in Patients receiving DCCR+Fenofibrate vs. Omega 3 Fatty Acids+Fenofibrate

Lipid data collected from both arms of the study of DCCR+fenofibrate (from baseline to day 126; described in 15.1), were compared with published lipid data from a double-blind placebo-controlled study in which prescription omega-3 fatty acids (p-OM3; Lovaza®) and fenofibrate and were co-administered to patients with very high triglyceride levels (>500 mg/dL; total of 128 men and women between the ages of 18 and 79 years of age)) (Roth et al., J Cardiovasc Pharmacol. 2009 September; 54(3):196-203; hereinafter “the Roth study”). All patients in the Roth study had a 6 week diet lead-in period, followed by random assignment to receive either p-OM3 (4 g/day) and fenofibrate (130 mg/day) or placebo (4 g/day) and fenofibrate (130 mg/day) for 8 weeks.

The median decrease in triglycerides, total cholesterol, non-HDL-C and VLDL-C for administration of fenofibrate and p-OM3 was similar to that of fenofibrate-only controls in both the Roth Study and the DCCR study. However, the median decrease observed for all of these lipid levels was much greater with DCCR and fenofibrate co-administration, thus showing a strong additive effect on the reduction of triglyceride levels.

The median increase in HDL-C was drastically higher for DCCR+fenofibrate vs. p-OM3+fenofibrate (about a 38% increase vs. about a 2% decrease, respectively). These data show that co-administration of DCCR and fenofibrate provide an substantial additive effect on increasing levels of HDL-C.

The median increase in levels of LDL-C were lower in the DCCR study compared with the Roth Study (about a 42% increase for DCCR+fenofibrate vs. about a 48% increase for p-OM3+fenofibrate). This reduced elevation of LDL-C with DCCR+fenofibrate treatment may provide a decreased risk of major cardiovacular events in these patients. The data from the Roth study and the study examining the combination of DCCR and fenofibrate are summarized in Table 53 below and in FIG. 29.

TABLE 53 Median Percentage Changes from Baseline for DCCR + fenofibrate vs. fenofibrate-only DCCR + Fenofibrate p-OM3 + Fenofibrate (Median Change) (Median Change) TG −77.8% −60.8% TC  −21% −8.0% Non-HDL-C −33.8% −8.2% VLDL-C −84.5% −57.6% LDL-C +41.5% +48.2% HDL-C +37.5 −1.9%

16. Pharmacokinetics of Diazoxide Choline in Humans

A pharmacokinetic study was conducted in 9 healthy male and female subjects who were between 18 to 75 years of age. In order to be included in the study, the subjects were required to have body mass index (BMI) between 22 and 35 kg/m², inclusive, and were required to weigh at least 50 kg. They had to be generally healthy as documented by the medical history, physical examination, vital sign assessments, 12-lead electrocardiogram (ECG), and clinical laboratory assessments. Their fasting triglyceride had to be ≧100 mg/dL and ≦1500 mg/dL, their fasting glucose ≦110 mg/dL and their HbAlc ≦6.0%.

Subjects were dosed for 10 days with a DCCR formulation. Each day they were administered 290 mg DCCR as two DCCR 145 mg tablets formulated as shown in Table 54. The study medication was taken orally, once daily, in the morning, within 15 minutes of a meal that included solid food. Pharmacokinetic samples were collected before initiation of treatment, and pre-dose after 2, 4, 6, and 8 days into the study. On the 10^(th) day of dosing, pharmacokinetic samples were collected pre-dose and 2, 4, 6, 8, 10, 12, 15 and 24 hours post-dosing.

TABLE 54 DCCR tablet composition for pharmacokinetic studies Component % wt Diazoxide choline 26.4%   PEO N750 30%  PEO 303 15%  Dibasic calcium phosphate 5% (Encompress) Talc 2% Colloidal silicon dioxide 1% Microcrystalline cellulose 19.1%   Magnesium Stearate 1.5%  Opadry white (tablet coating) 3%

Steady state circulating drug levels were observed between 8 and 9 days after the start of treatment. Circulating diazoxide concentrations were measured using a sensitive, precise, reproducible, validated assay. The following pharmacokinetic variables were measured on study day 10 (representing steady state): AUC_(0-24(ss)), the area under the plasma concentration-time curve from time zero to 24 hours at steady state, using the linear trapezoidal rule; C_(max(ss)), the maximum or peak measured plasma concentration at steady state; C_(min(ss)), the minimum or trough plasma concentration at steady state; C_(av(ss)), the average plasma concentration at steady state obtained by dividing AUC_(0-24(ss)) by 24; Fluctuation Index, the fluctuation at steady state calculated as [(C_(max(ss))−C_(min(ss))/C_(av(ss))]; and % Peak-to-Trough Fluctuation, the Fluctuation Index×100. The collected data are summarized in Table 55 below.

TABLE 55 Pharmacokinetic Data for Patients Parameter Range Mean ± SD Median AUC_(0-24(ss)) (μg*hr/mL) 344.5-750.7  546.0 ± 128.4 542.6 C_(max(ss)) (μg/mL) 16.3-35.4 25.9 ± 5.8 26.6 C_(min(ss)) (μg/mL) 10.4-28.9 19.6 ± 5.3 18.4 C_(av(ss)) (μg/mL) 14.4-31.3 22.8 ± 5.3 22.6 % Peak-to-Trough Fluctuation 18.0%-41.1%  28.5 ± 7.2% 27.8% 17. Co-administration of Diazoxide Choline and Fenofibrate Choline in Patients with Very High Triglycerides

A group of 4 patients from the study described in Example 15, all males ranging in age from 28 to 62 years old, with very high triglycerides and low HDL were treated with 290 mg DCCR for 12 weeks and with both 290 mg DCCR and 135 mg fenofibrate for an additional 2 weeks. All subjects at baseline had triglycerides above 500 mg/dL (very high triglycerides) and HDL-C below 40 mg/dL (low HDL-C). Additionally, all four patients had elevated non-HDL-C. The patients were at elevated risk of cardiovascular events due to marked elevations of non-HDL-C and low HDL-C.

Combined treatment with DCCR and fenofibrate in these patients normalized triglycerides (TG <150 mg/dL), reduced non-HDL-C by 50%, and normalized HDL-C (increased by 58.5%). The risk of cardiovascular events in these patients was reduced significantly by treatment with DCCR and fenofibrate. Table 56 below summarizes the data averages for the lipid characteristics of each patient at baseline and end of treatment.

TABLE 56 Average Baseline and End-of-Treatment Lipid Parameters End of Baseline Treatment Change % Change Triglycerides (mg/dL) 728.8 115.5 −613.3 −84.20% Non-HDL-C (mg/dL) 301.7 151 −150.7 −49.90% HDL-C (mg/dL) 29.5 46.8 17.3 58.50%

18. Characterizing Bioavailability of Diazoxide Choline in Mice

A study was conducted in male mice to characterize the absolute bioavailability of diazoxide choline polymorph form B and the relative bioavailability of four structurally related molecules, all of which agonize the K_(ATP) channel. The related molecules have the same ring structure as diazoxide, namely 4H-1,2,4-benzothiadiazine 1,1-dioxide, but differ in the constituent at positions 3, 6 and 7. Position 6 was occupied either by hydrogen, chlorine, bromine, or fluorine. Position 7 was occupied either by hydrogen, fluorine, or methoxy. The substitutions at the 3 position included 3-amino-isopropylene and 3-amino-cyclobutane.

The study used 4 mice dosed per treatment arm. Following a single intravenous or oral dose, administered by gavage, pharmacokinetic samples were collected at 4 time points: 0.5, 1, 3, and 6 hours post-dosing. Two mice were sampled at each timepoint, with each mouse contributing samples to two timepoints. Each test drug was administered as a 50 mg/kg dose, which yields nearly equal molar concentrations. Diazoxide choline was administered as a diazoxide equivalent dose of 50 mg/kg. Blood samples were collected into tubes containing dipotassium EDTA as the anticoagulant and placed on ice packs until centrifugation (at 3000 rpm for 10 minutes at 4° C.). After centrifugation, samples were stored at −70° C. until they were thawed for analysis. Circulating drug levels were measured using a precise and reproducible LC-MS/MS assay.

The following pharmacokinetic parameters were measured: AUC₀₋₆, the area under the plasma concentration versus time curve from time 0 to 6 hours post-dosing calculated by the linear trapezoidal method; and C_(max), the maximum measured plasma concentration over the time from 0 to 6 hours post-dosing. Absolute oral bioavailability of diazoxide choline was calculated by comparing the AUC₀₋₆ of the oral dose to that of the IV dose. Relative oral bioavailability of the remaining KATP channel agonists was calculated as the ratio of the AUC₀₋₆ for the molecule to that of the IV dose of diazoxide choline. A summary of the data obtained from the mouse study are summarized in Table 57 as follows.

TABLE 57 Bioavailability of DCCR and Related Molecules in Male Mice Absolute Relative Mode of AUC₀₋₆ C_(max) oral oral Molecule admin. (ng*hr/ml) (ng/mL) bioavailability bioavail. Diazoxide IV 137176 ± 3352  34310 ± 3671 — — choline Diazoxide Oral 141260 ± 24134  29292 ± 2685 100% — choline gavage 7H-6-methoxy- Oral 3590 ± 1860 1560 ± 982 — 2.6% 3-amino- gavage isopropylene- 4H-1,2,4- benzothiadiazine 1,1-dioxide 7-chloro-6- Oral 313 ± 15   80 ± 12 — 0.2% fluoro-3- gavage amino- isopropylene- 4H-1,2,4- benzothiadiazine 1,1-dioxide 7-bromo-6H-- Oral 800 ± 126 181 ± 16 — 0.6% 3-amino- gavage cyclobutane- 4H-1,2,4- benzothiadiazine 1,1-dioxide 7-fluoro-6H-3- Oral 89 ± 16  24 ± 10 — 0.06%  amino- gavage cyclobutane- 4H-1,2,4- benzothiadiazine 1,1-dioxide

The use of the choline salt of diazoxide provides for complete oral bioavailability of diazoxide (100%) in this mouse study, while administration of the free base of a range of structurally related molecules results in very limited oral bioavailability, between 0.06% and 2.6%.

19. Intravenous Administration of DCCR For Patients with Acute Pancreatitis

A patient with acute pancreatitis is given a bolus dose of diazoxide choline (between about 0.5 to 5 mg/kg) combined with insulin (typically in the range of 50-200 mg) over the course of 5-10 minutes, followed by continuous administration of diazoxide choline at a rate of about 0.02 to 0.2 mg/kg/hr to a total daily dosage of between about 100-400 mg/day, up to 500 mg/day. Insulin is co-administered at a rate of that is sufficient to maintain fasting glucose between about 70 mg/dL and 125 mg/dL and ensure peak post prandial glucose is less than 200 mg/dL. This may be accomplished using a basal insulin, a fast-acting prandial insulin, or both in combination. Administration of the combination of diazoxide choline and insulin may be continued until the pancreatitis has subsided.

All patents and other references cited in the specification are indicative of the level of skill of those skilled in the art to which the invention pertains, and are incorporated by reference in their entireties, including any tables and figures, to the same extent as if each reference had been incorporated by reference in its entirety individually.

One skilled in the art would readily appreciate that the present invention is well adapted to obtain the ends and advantages mentioned, as well as those inherent therein. The methods, variances, and compositions described herein as presently representative of preferred embodiments are exemplary and are not intended as limitations on the scope. Changes therein and other uses will occur to those skilled in the art, which are encompassed within the spirit of the invention, are defined by the scope of the claims.

Definitions provided herein are not intended to be limiting from the meaning commonly understood by one of skill in the art unless indicated otherwise.

The inventions illustratively described herein may suitably be practiced in the absence of any element or elements, limitation or limitations, not specifically disclosed herein. Thus, for example, the terms “comprising”, “including,” containing”, etc. shall be read expansively and without limitation. Additionally, the terms and expressions employed herein have been used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the invention claimed. Thus, it should be understood that although the present invention has been specifically disclosed by preferred embodiments and optional features, modification and variation of the inventions embodied therein herein disclosed may be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of this invention.

The invention has been described broadly and generically herein. Each of the narrower species and subgeneric groupings falling within the generic disclosure also form part of the invention. This includes the generic description of the invention with a proviso or negative limitation removing any subject matter from the genus, regardless of whether or not the excised material is specifically recited herein. Other embodiments are within the following claims. In addition, where features or aspects of the invention are described in terms of Markush groups, those skilled in the art will recognize that the invention is also thereby described in terms of any individual member or subgroup of members of the Markush group. 

That which is claimed is:
 1. A method for treating dyslipidemia in a subject, the method comprising administering to the subject: a. a formulation comprising one or more omega-3 fatty acids, and b. a formulation comprising a therapeutically effective amount of a K_(ATP) channel opener, said K_(ATP) channel opener comprising: (i) an anion of a K_(ATP) channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV,

wherein in Formulas I and II: R¹ is selected from the group consisting of hydrogen, C₁-C₆ alkyl, substituted C₁-C₆ alkyl, cycloalkyl, and substituted cycloalkyl provided however that when R¹ is a substituted C₁-C₆ alkyl or a substituted cycloalkyl, then the substituent does not include an amino group; R^(2a) is hydrogen; R^(2b) is hydrogen; X is a 1, 2 or 3 atom chain, wherein each atom is independently selected from carbon, sulfur and nitrogen, and each atom is optionally substituted with halogen, hydroxyl, C₁-C₆ alkyl, substituted C₁-C₆ alkyl, C₁-C₆ alkoxy, cycloalkyl, substituted cycloalkyl, or substituted C₁-C₆ alkoxy, provided however that when an atom of the chain is substituted with substituted C₁-C₆ alkyl, substituted C₁-C₆ alkoxy or substituted cycloalkyl, then the substituent does not include an amino group; and ring B is saturated, monounsaturated, polyunsaturated or aromatic, and wherein in Formulas III and IV: R¹ is selected from the group consisting of hydrogen, C₁-C₆ alkyl, substituted C₁-C₆ alkyl, and cycloalkyl provided however that when R¹ is a substituted C₁-C₆ alkyl, then the substituent does not include an amino group; R^(2a) is hydrogen; R^(2b) is hydrogen; R³ is selected from the group consisting of hydrogen, halogen, C₁-C₆ alkyl, substituted C₁-C₆ alkyl, cycloalkyl and substituted cycloalkyl provided however that when R³ is a substituted C₁-C₆ alkyl, then the substituent does not include an amino group; R⁴ is selected from the group consisting of hydrogen, halogen, C₁-C₆ alkyl, substituted C₁-C₆ alkyl, cycloalkyl and substituted cycloalkyl provided however that when R⁴ is a substituted C₁-C₆ alkyl, then the substituent does not include an amino group, and (ii) a cation selected from the group consisting of an alkali metal and a compound comprising an ammonium group comprising at least one tertiary amine group.
 2. The method of claim 1, wherein said dyslipidemia is selected from the group consisting of abnormally high total cholesterol level in a subject's blood, abnormally high LDL cholesterol, abnormally high VLDL cholesterol, abnormally high non-HDL cholesterol, abnormally high triglycerides, abnormally low HDL cholesterol, or any combination thereof.
 3. The method of claim 1, wherein said dyslipidemia comprises abnormally high triglycerides.
 4. The method of claim 3, wherein said abnormally high triglycerides comprises a triglyceride level of at least about 500 mg/dL.
 5. The method of claim 1, wherein said K_(ATP) channel opener is formulated as a controlled release oral formulation.
 6. The method of claim 5, wherein said controlled release oral formulation is formulated for once or twice daily administration.
 7. The method of claim 1, wherein said subject is administered about 145 to 435 mg per day of said K_(ATP) channel opener.
 8. The method of claim 1, wherein said K_(ATP) channel opener comprises a salt of diazoxide.
 9. The method of claim 1, wherein said K_(ATP) channel opener comprises diazoxide choline.
 10. The method of claim 1, wherein said omega-3 fatty acid formulation is substantially free of DHA.
 11. A method for initiating treatment for a dyslipidemia in a subject, the method comprising administering a fibrate or a salt thereof and a K_(ATP) channel opener, said K_(ATP) channel opener comprising: (i) an anion of a K_(ATP) channel opener selected from the group consisting of Formula I, Formula II, Formula III and Formula IV,

wherein in Formulas I and II: R¹ is selected from the group consisting of hydrogen, C₁-C₆ alkyl, substituted C₁-C₆ alkyl, cycloalkyl, and substituted cycloalkyl provided however that when R¹ is a substituted C₁-C₆ alkyl or a substituted cycloalkyl, then the substituent does not include an amino group; R^(2a) is hydrogen; R^(2b) is hydrogen; X is a 1, 2 or 3 atom chain, wherein each atom is independently selected from carbon, sulfur and nitrogen, and each atom is optionally substituted with halogen, hydroxyl, C₁-C₆ alkyl, substituted C₁-C₆ alkyl, C₁-C₆ alkoxy, cycloalkyl, substituted cycloalkyl, or substituted C₁-C₆ alkoxy, provided however that when an atom of the chain is substituted with substituted C₁-C₆ alkyl, substituted C₁-C₆ alkoxy or substituted cycloalkyl, then the substituent does not include an amino group; and ring B is saturated, monounsaturated, polyunsaturated or aromatic, and wherein in Formulas III and IV: R¹ is selected from the group consisting of hydrogen, C₁-C₆ alkyl, substituted C₁-C₆ alkyl, and cycloalkyl provided however that when R¹ is a substituted C₁-C₆ alkyl, then the substituent does not include an amino group; R^(2a) is hydrogen; R^(2b) is hydrogen; R³ is selected from the group consisting of hydrogen, halogen, C₁-C₆ alkyl, substituted C₁-C₆ alkyl, cycloalkyl and substituted cycloalkyl provided however that when R³ is a substituted C₁-C₆ alkyl, then the substituent does not include an amino group; R⁴ is selected from the group consisting of hydrogen, halogen, C₁-C₆ alkyl, substituted C₁-C₆ alkyl, cycloalkyl and substituted cycloalkyl provided however that when R⁴ is a substituted C₁-C₆ alkyl, then the substituent does not include an amino group, and (ii) a cation selected from the group consisting of an alkali metal and a compound comprising an ammonium group comprising at least one tertiary amine group; wherein said fibrate or a salt thereof and said K_(ATP) channel opener are both initially administered at sub-optimal titration doses, and the doses of both are increased to maintenance doses over 7 to 28 days.
 12. The method of claim 11, wherein said dyslipidemia comprises a triglyceride level of at least about 500 mg/dL and/or a HDL-C level of less than about 40 mg/dL.
 13. The method of claim 11, wherein said K_(ATP) channel opener comprises a salt of diazoxide.
 14. The method of claim 11, wherein said K_(ATP) channel opener comprises diazoxide choline.
 15. The method of claim 11, wherein said fibrate or a salt thereof comprises fenofibrate or a salt thereof.
 16. The method of claim 11, wherein said fibrate or a salt thereof comprises fenofibrate choline.
 17. A method for treating a subject with acute pancreatitis, the method comprising intravenously administering a bolus dose of a K_(ATP) channel opener of between about 0.5 to 5 mg/kg over 5 to 10 minutes; wherein said K_(ATP) channel opener is selected from the group consisting of Formula I, Formula II, Formula III and Formula IV,

wherein in Formulas I and II: R¹ is selected from the group consisting of hydrogen, C₁-C₆ alkyl, substituted C₁-C₆ alkyl, cycloalkyl, and substituted cycloalkyl provided however that when R¹ is a substituted C₁-C₆ alkyl or a substituted cycloalkyl, then the substituent does not include an amino group; R^(2a) is hydrogen; R^(2b) is hydrogen; X is a 1, 2 or 3 atom chain, wherein each atom is independently selected from carbon, sulfur and nitrogen, and each atom is optionally substituted with halogen, hydroxyl, C₁-C₆ alkyl, substituted C₁-C₆ alkyl, C₁-C₆ alkoxy, cycloalkyl, substituted cycloalkyl, or substituted C₁-C₆ alkoxy, provided however that when an atom of the chain is substituted with substituted C₁-C₆ alkyl, substituted C₁-C₆ alkoxy or substituted cycloalkyl, then the substituent does not include an amino group; and ring B is saturated, monounsaturated, polyunsaturated or aromatic, and wherein in Formulas III and IV: R¹ is selected from the group consisting of hydrogen, C₁-C₆ alkyl, substituted C₁-C₆ alkyl, and cycloalkyl provided however that when R¹ is a substituted C₁-C₆ alkyl, then the substituent does not include an amino group; R^(2a) is hydrogen; R^(2b) is hydrogen; R³ is selected from the group consisting of hydrogen, halogen, C₁-C₆ alkyl, substituted C₁-C₆ alkyl, cycloalkyl and substituted cycloalkyl provided however that when R³ is a substituted C₁-C₆ alkyl, then the substituent does not include an amino group; R⁴ is selected from the group consisting of hydrogen, halogen, C₁-C₆ alkyl, substituted C₁-C₆ alkyl, cycloalkyl and substituted cycloalkyl provided however that when R⁴ is a substituted C₁-C₆ alkyl, then the substituent does not include an amino group. and said K_(ATP) channel opener is formulated for intravenous administration.
 18. The method of claim 17, further comprising additional administration of the K_(ATP) channel opener after completion of the bolus at between about 0.02 to 0.2 mg/kg/hr, along with administration of insulin on a recurring basis at a level and rate sufficient to maintain fasting glucose between about 70 mg/dL and 125 mg/dL and peak post prandial glucose less than 200 mg/dL.
 19. The method of claim 17, wherein the K_(ATP) channel opener comprises diazoxide or an ion thereof.
 20. The method of claim 17, further comprising co-administration of nicorandil. 